CN111601613A - Methods of treating metabolic disorders with FGF21 variants - Google Patents

Abstract

Provided herein are methods of treating, preventing and managing metabolic or cardiovascular disorders, and methods of reducing cardiovascular risk using FGF21 protein variants, including Fc-FGF21 variant fusion proteins.

Description

Methods of treating metabolic disorders with FGF21 variants

I. Technical Field

The present disclosure relates to methods of treating, preventing and managing metabolic or cardiovascular disorders, and to methods of reducing cardiovascular risk with FGF21 protein variants, including Fc-FGF21 variant fusion proteins.

II. Background Art

Insulin resistance is common and plays an important role in the pathogenesis of a variety of diseases. Central adiposity is an important source of inflammatory cytokines and non-esterified fatty acids, which have been shown to have adverse effects on insulin sensitivity. Metabolic disorders associated with insulin resistance and hyperlipidemia manifest in various forms, such as diabetes, obesity, dyslipidemia, coronary heart disease, and NAFLD/NASH. Fibroblast growth factor 21 (FGF21) is thought to act on liver and adipose tissue to improve insulin sensitivity, lower triglycerides, and reduce hyperlipidemia. Therefore, FGF21 analogs with clinically acceptable dosing regimens are expected to be front-line therapies for these metabolic disorders.

FGF21 is a member of the fibroblast growth factor family and has diverse metabolic functions. FGF21 is released primarily from hepatocytes, and to a lesser extent from adipose and pancreatic beta cells. In mice, FGF21 has been shown to be a key regulator of metabolism in which expression is induced under conditions of both starvation and obesity. FGF21 induces various beneficial metabolic effects, including weight loss, improved insulin sensitivity, increased high-density lipoprotein-cholesterol (HDL-C), and decreased Low-density lipoprotein-cholesterol (LDL-C), triglyceride (TG) and liver TG content. Furthermore, in a nonclinical animal model of diabetes, FGF21 effectively lowered glucose without inducing hypoglycemia. In T2DM patients with increased body mass index, four weeks of treatment with FGF21 protein analogs resulted in significant improvements in lipid profiles, indicating the translatability of the metabolic effects observed in animal models to humans (Gaich et al., 2013 , Cell Metab. [Cell Metabolism] 18(3):333-40; Dong et al., 2015, Br J ClinPharmacol. [British Journal of Clinical Pharmacology] doi:10.1111/bcp.12676). A randomized, placebo-controlled, double-blind proof-of-concept trial by Gaich et al in patients with obesity and type 2 diabetes (where patients received placebo or LY2405319 at 3, 10, or 20 mg daily for 28 days) LY2405319 (a variant of FGF21) is described in . Dong et al. describe a first-in-human study to evaluate the pharmacokinetics/pharmacodynamics of a single intravenous (IV) dose of PF-05231023, a long-acting fibroblast growth factor 21 (FGF21) analog PK/PD, safety and tolerability, and reported that PF-05231023 levels peaked immediately after IV dosing, with a mean terminal half-life of 6.5h- for intact C- and N-termini, respectively 7.7h and 66.5h-96.6h.

There remains a need for improved FGF21 therapy with clinically acceptable dosing regimens for treating metabolic disease and reducing cardiovascular risk in patients.

III. SUMMARY OF THE INVENTION

Novel modifications to FGF21 can improve half-life and/or potency compared to wild-type FGF21 to provide for the treatment or management of metabolic or cardiovascular disorders (e.g., one or more alleviating disorders) with clinically preferred dosing regimens. various symptoms) or FGF21 therapeutics for cardiovascular risk reduction.

In particular aspects, provided herein are FGF21 protein variants (eg, FGF21 protein variants described in Table 1, eg, Fc-fusion proteins) and pharmaceutical compositions comprising such FGF21 protein variants for use in treatment, prevention and/or Or manage hypertriglyceridemia and heart disease risk, insulin resistance (eg, in patients with genetic mutations in the insulin receptor) and lipodystrophy, diabetes, obesity, and nonalcoholic fatty liver disease (NAFLD)/nonalcoholic Methods of Steatohepatitis (NASH).

V103 is a genetically engineered variant of human FGF21 stabilized via the introduction of a new disulfide bond and fused at its N-terminus to human IgGl Fc (see PCT Publication WO 2013/049247, which is hereby incorporated by reference in its entirety) ). The combination of stability and fusion of the human FGF21 mutant with IgGl Fc resulted in an approximately 50-100-fold increase in half-life in cynomolgus monkeys (5 to 8 days) compared to wild-type (WT) human FGF21 (approximately 2 hours).

V103 replicates the published effects of FGF21 in an animal model of insulin-resistant T2DM, including reductions in body weight, glucose, insulin, serum TG, and hepatic TG levels. V103 also showed significant reductions in liver lipids, fibrosis and inflammation in a mouse model of nonalcoholic steatohepatitis (NASH). Accordingly, provided herein are pharmaceutical compositions comprising FGF21 protein variant V103 (SEQ ID NO: 11) for the treatment, prevention and/or management of metabolic disorders (including hypertriglyceridemia and heart disease risk, insulin resistance (e.g. patients with genetic mutations of the insulin receptor) and lipodystrophy, diabetes, obesity, and nonalcoholic fatty liver disease (NAFLD)/nonalcoholic steatohepatitis (NASH).

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing a metabolic disorder or a cardiovascular disorder in a human subject, wherein the human FGF21 protein variant is administered in an amount of 100 mg to 600 mg Doses within the range are provided for administration. In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing a metabolic disorder or a cardiovascular disorder in a human subject, wherein the human FGF21 protein variant is administered in an amount of 100 mg to 600 mg Amounts within the range are provided for administration.

In some embodiments of this aspect, the metabolic disorder or cardiovascular disorder is selected from the group consisting of hypercholesterolemia, dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH) ), type 2 diabetes and obesity.

In some embodiments of this aspect, treating, preventing or managing a metabolic or cardiovascular disorder comprises or is characterized by reducing one or more of the following: a subject's body weight, liver fat content, elevated LDL-C, Total C, triglycerides and Apo B levels. In certain embodiments thereof, treating, preventing or managing a metabolic disorder or cardiovascular disorder comprises or is characterized by increasing HDL-C levels in the subject. In some embodiments thereof, treating, preventing or managing a metabolic or cardiovascular disorder comprises or is characterized by reducing the subject's triglyceride levels by at least about 40% or at least about 50%. In some embodiments thereof, treating, preventing or managing a metabolic or cardiovascular disorder comprises or is characterized by reducing cardiovascular risk in a subject.

In some embodiments of this aspect, the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of : Q55C, R105K, G148C, K150R, P158S, S195A, P199G and G202A numbered according to SEQ ID NO:1. In certain embodiments thereof, the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In some embodiments of this aspect, the human FGF21 protein variant is provided for administration in a dose of at least 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, or 400 mg. In some embodiments thereof, the human FGF21 protein variant is provided for administration at a dose of about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, or 190 mg, optionally wherein the human FGF21 protein Variants are provided in doses of approximately 100 mg or 150 mg. In some embodiments thereof, the human FGF21 protein variant is provided at a dose of about 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg For administration, optionally wherein the human FGF21 protein variant is provided in a dose of about 200 mg, 250 mg, or 300 mg.

In some embodiments of this aspect, the human FGF21 protein variant is provided for subcutaneous administration. In some embodiments, the human FGF21 protein variant is provided for administration once a month or once every 4 weeks, once every 3 weeks, once every 2 weeks, or once a week. In some embodiments thereof, the human FGF21 protein variant is provided for administration once a month or once every 4 weeks, once every 3 weeks, once every 2 weeks or once a week. In some embodiments thereof, the human FGF21 protein variant is provided for subcutaneous administration.

In one aspect, provided herein is a human FGF21 protein variant in the manufacture of a medicament for the treatment, prevention or management of a metabolic disorder or cardiovascular disorder, wherein the unit dose of the human FGF21 protein variant is in the range of 100 mg to 600 mg . In one aspect, provided herein is a human FGF21 protein variant in the manufacture of a medicament for the treatment, prevention or management of a metabolic or cardiovascular disorder, wherein the amount of the human FGF21 protein variant is in the range of 100 mg to 600 mg.

In some embodiments of this aspect, the metabolic disorder or cardiovascular disorder is selected from the group consisting of hypercholesterolemia, dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH) ), type 2 diabetes and obesity.

In some embodiments of this aspect, treating, preventing or managing a metabolic or cardiovascular disorder comprises or is characterized by a reduction in one or more of the following: body weight, liver fat content, elevated LDL-C, total C, glycerol Triester and Apo B levels. In some embodiments thereof, the treatment, prevention or management of a metabolic or cardiovascular disorder comprises or is characterized by increasing HDL-C levels. In some embodiments thereof, triglyceride levels are reduced by at least about 40% or at least about 50%. In some embodiments thereof, treating, preventing or managing a metabolic or cardiovascular disorder comprises or is characterized by reducing cardiovascular risk.

In some embodiments of this aspect, the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of : Q55C, R105K, G148C, K150R, P158S, S195A, P199G and G202A numbered according to SEQ ID NO:1. In some embodiments thereof, the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In some embodiments of this aspect, the human FGF21 protein variant is provided for administration in a dose of at least 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, or 400 mg. In some embodiments, the human FGF21 protein variant is provided for administration at a dose of about 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, or 400 mg. In some embodiments, the human FGF21 protein variant is provided for administration at a dose of about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, or 190 mg, optionally wherein the human FGF21 protein is mutated The body is provided for administration in doses of approximately 100 mg or 150 mg. In some embodiments, the human FGF21 protein variant is provided at a dose of about 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg For administration, optionally wherein the human FGF21 protein variant is provided for administration in a dose of about 200 mg, 250 mg or 300 mg.

In some embodiments, the human FGF21 protein variant is provided for administration once a month or once every 4 weeks, once every 3 weeks, once every 2 weeks, or once a week. In some embodiments, the human FGF21 protein variant is provided for administration once a month or once every 4 weeks, once every 3 weeks, once every 2 weeks, or once a week. In some embodiments, the human FGF21 protein variant is formulated for subcutaneous administration. In some embodiments, the human FGF21 protein variant is provided for subcutaneous administration.

In one aspect, provided herein is a method of treating, preventing or managing a metabolic disorder or a cardiovascular disorder in a human subject, wherein the method comprises administering to the subject a human FGF21 protein variant at a dose in the range of 100 mg to 600 mg . In one aspect, provided herein is a method of treating, preventing or managing a metabolic disorder or a cardiovascular disorder in a human subject, wherein the method comprises administering to the subject a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg .

In some embodiments, the metabolic disorder or cardiovascular disorder is selected from hypercholesterolemia, dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), 2 type diabetes and obesity.

In some embodiments, treating, preventing or managing a metabolic or cardiovascular disorder comprises or is characterized by reducing one or more of the following: a subject's body weight, liver fat content, elevated LDL-C, total C, Triglyceride and Apo B levels. In certain embodiments, treating, preventing or managing a metabolic disorder or cardiovascular disorder comprises or is characterized by increasing HDL-C levels in the subject. In some embodiments, treating, preventing or managing a metabolic or cardiovascular disorder comprises or is characterized by reducing the subject's triglyceride levels by at least about 40% or at least about 50%. In some embodiments, treating, preventing or managing a metabolic or cardiovascular disorder comprises or is characterized by reducing a subject's cardiovascular risk.

In some embodiments, the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of: according to SEQ IDNO: 1 numbered Q55C, R105K, G148C, K150R, P158S, S195A, P199G and G202A. In certain embodiments, the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In some embodiments, the method comprises administering to the subject the human FGF21 protein variant at a dose of at least 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, or 400 mg. In some embodiments, the method comprises administering to the subject the human FGF21 protein variant at a dose of about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, or 190 mg. In some embodiments, the method comprises administering to the subject the human FGF21 protein variant at a dose of about 100 mg or 150 mg. In some embodiments, the method comprises administering the human to the subject at a dose of about 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg FGF21 protein variants. In some embodiments, the method comprises administering to the subject the human FGF21 protein variant at a dose of about 200 mg, 250 mg, or 300 mg.

In some embodiments, the human FGF21 protein variant is administered once a month or once every 4 weeks, once every 3 weeks, once every 2 weeks, or once a week. In some embodiments, the human FGF21 protein variant is administered once a month or once every 4 weeks, once every 3 weeks, once every 2 weeks, or once a week. In some embodiments, the method comprises administering the human FGF21 protein variant in a form suitable for subcutaneous administration. In some embodiments, the method comprises subcutaneously administering the human FGF21 protein variant. In some embodiments, the human FGF21 protein variant is administered subcutaneously.

Each of the foregoing aspects and embodiments, as well as other elements described herein, may be combined in any manner and without limitation.

IV. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

Provided herein are methods and pharmaceutical compositions for use in the treatment, prevention, or management of metabolic or cardiovascular disorders (eg, alleviating one or more symptoms of the disorder), including in doses or amounts in the range of 100 mg to 600 mg A human FGF21 protein variant (eg, an Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)) is administered to a subject in need thereof. In particular aspects, the human FGF21 protein variant (eg, V103 (SEQ ID NO: 11)) is administered at a dose ranging from 200 mg to 400 mg. In particular aspects, the human FGF21 protein variant (eg, V103 (SEQ ID NO: 11)) is administered at a dose of at least 200 mg, at least 300 mg, or at least 400 mg. In particular aspects, the human FGF21 protein variant (eg, an Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)) is administered every 4 weeks at a dose ranging from 100 mg to 600 mg (eg, 250 mg to 350 mg) (or monthly), or every 3 weeks or every 2 weeks.

In one aspect, non-limiting examples of metabolic or cardiovascular disorders to be treated, prevented or managed by the methods provided herein include hypertriglyceridemia, diabetes (eg, type 2 diabetes), obesity, type 1 diabetes, Pancreatitis, dyslipidemia, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), insulin resistance, hyperinsulinemia, glucose intolerance, hyperglycemia, metabolic syndrome, hypertension, Cardiovascular disease, acute myocardial infarction, atherosclerosis, peripheral arterial disease, stroke, heart failure, coronary heart disease, kidney disease, diabetic complications, neuropathy, diseases associated with severely inactivating mutations in the insulin receptor and/or gastric paralysis.

the term

With respect to FGF21, the term "native" or "wild-type" refers to biologically active naturally occurring FGF21, including biologically active naturally occurring FGF21 variants. An exemplary human FGF21 wild-type sequence has NCBI reference number NP_061986.1 and can be found in such issued patents, eg, US 6,716,626B1 (SEQ ID NO: 1).

The corresponding mRNA sequence encoding the full-length FGF21 polypeptide (NCBI reference number NM_019113.2) is shown below (SEQ ID NO: 2):

The mature FGF21 sequence lacks a leader sequence and may also include other modifications of the polypeptide, such as proteolytic processing of the amino terminus (with or without a leader sequence) and/or carboxy terminus, cleavage of smaller polypeptides from larger precursors, N- Linked and/or O-linked glycosylation, and other post-translational modifications understood by those skilled in the art. A representative example of a mature FGF21 sequence has the following sequence (SEQ ID NO: 3, which represents amino acid positions 29-209 of the full-length FGF21 protein sequence (NCBI reference number NP_061986.1)):

The corresponding cDNA sequence encoding the mature FGF21 polypeptide (SEQ ID NO:3) is shown below (SEQ ID NO:4):

The term "FGF21 protein variant", "human FGF21 variant", "FGF21 polypeptide or protein variant", "FGF21 mutant" or any similar term is defined as comprising the amino acid sequence of FGF21 in which the naturally occurring (ie wild-type) amino acid sequence has been Human FGF21 that is modified (eg, wherein at least one amino acid of the wild-type protein has been substituted with another amino acid and/or removed). Additionally, variants may include N- and/or C-terminal truncations relative to wild-type FGF21 protein, or internal amino acid deletions relative to wild-type human FGF21 protein. Generally, a variant has some modified property, structure or function of the wild-type protein. For example, a variant may have enhanced or improved physical stability in concentrated solutions (eg, less hydrophobicity-mediated aggregation), enhanced or improved plasma stability when incubated with plasma, maintained favorable biological activity Enhanced or improved biological activity at the same time.

Acceptable amino acid substitutions and modifications that constitute the differences between the FGF21 polypeptides and protein variants and mutants of the methods provided herein and wild-type FGF21 include, but are not limited to, one or more amino acid substitutions (including analogs with non-naturally occurring amino acids) substitution) and truncation. Thus, FGF21 protein variants include, but are not limited to, site-directed FGF21 mutants, truncated FGF21 polypeptides, proteolysis-resistant FGF21 mutants, aggregation-reducing FGF21 mutants, FGF21 combination mutants, FGF21 conjugates as described herein (eg, fatty acid-FGF21 conjugates, PEG-FGF21 conjugates) and FGF21 fusion proteins (eg, Fc domain fusion proteins, human serum albumin fusion proteins).

The variants may have increased compatibility with pharmaceutical preservatives (eg m-cresol, phenol, benzyl alcohol), thus enabling the preparation of preserved pharmaceutical formulations that maintain the physiochemical properties and biological activity of the protein during storage. Thus, variants with enhanced drug stability relative to wild-type FGF21 have improved physical stability in concentrated solutions under physiological and stored drug formulation conditions, while maintaining biological efficacy. As a set of non-limiting examples, the variants provided herein can be more resistant to proteolytic and enzymatic degradation; can have improved stability; and are less likely to aggregate compared to their wild-type counterparts. As used herein, these terms are not mutually exclusive or limiting, and it is entirely possible that a given variant has one or more modified properties of the wild-type protein.

In particular aspects, the FGF21 protein variant is a biologically active variant of a wild-type FGF21 protein that exhibits one or more biological activities of wild-type FGF21. The previously reported biological activities of wild-type FGF21 include, but are not limited to, the following: (i) FGF21 has been shown to induce insulin-independent glucose uptake; (ii) FGF21 has also been shown to ameliorate hyperglycemia in a range of diabetic rodent models; (iii) FGF21-overexpressing transgenic mice were found to be resistant to diet-induced metabolic abnormalities and displayed reduced body weight and fat mass, as well as enhanced insulin sensitivity (Badman, M.K. et al. (2007) Cell Metab 5,426 -37);(iv) Administration of FGF21 to diabetic non-human primates resulted in a decrease in fasting blood glucose, triglycerides, insulin and glucagon levels and in a significant improvement in lipoprotein profiles, including a nearly 80% increase in HDL cholesterol (Kharitonenkov, A. et al., 2007, Endocrinology 148, 774-81); (v) FGF21 acts as an important endocrine hormone that helps control adaptation to the fasting state (Badman et al. 2009, Endocrinology [Endocrinology] 150, 4931; Inagaki et al., 2007, Cell Metabolism [Cell Metabolism] 5, 415); and (vi) FGF21 can modulate downstream markers including but not limited to glucose or 2-deoxy-glucose uptake, pERK and other phosphorylation or acetylation protein or NAD levels.

The term "native Fc" refers to a molecule or sequence comprising a non-antigen-binding fragment sequence resulting from digestion of an intact antibody or otherwise produced, in monomeric or multimeric form, and which may contain a hinge region. The original immunoglobulin source for native Fc is preferably of human origin and can be any immunoglobulin, although IgGl and IgG2 are preferred. Native Fc molecules are composed of monomeric polypeptides that can be linked into dimeric or multimeric forms by covalent (ie, disulfide bonds) and non-covalent associations. The number of intermolecular disulfide bonds between monomeric subunits of native Fc molecules ranges from 1 to 4, depending on the class (eg, IgG, IgA, and IgE) or subclass (eg, IgG1, IgG2, IgG3, IgA1, and IgGA2). An example of a native Fc is the disulfide-bonded dimer produced by papain digestion of IgG (see Ellison et al., 1982, Nucleic Acids Res. [Nucleic Acids Res.] 10:4071-9). The term "native Fc" as used herein is generic for monomeric, dimeric and multimeric forms.

The term "Fc variant" refers to a molecule or sequence that is modified from a native Fc but still contains a binding site for the salvage receptor FcRn (the neonatal Fc receptor). International Publication Nos. WO 97/34631 and WO 96/32478 describe exemplary Fc variants, and interactions with salvage receptors, and are hereby incorporated by reference for this purpose. Thus, the term "Fc variant" may encompass humanized molecules or sequences from non-human native Fc. In addition, native Fcs contain regions that can be removed because they provide structural features or biological activities that are not required for the fusion molecules of the fusion proteins of the invention. Thus, the term "Fc variant" encompasses molecules or sequences that lack, or are modified in, one or more native Fc sites or residues that affect or Involved in: (1) disulfide bond formation, (2) incompatibility with selected host cells, (3) N-terminal heterogeneity after expression in selected host cells, (4) glycosylation, (5) ) interaction with complement, (6) binding to Fc receptors other than salvage receptors, and/or (7) antibody-dependent cellular cytotoxicity (ADCC). Fc variants are described in further detail below.

The term "Fc domain" encompasses native Fc as defined above, as well as Fc variants and sequences. As with Fc variants and native Fc molecules, the term "Fc domain" includes molecules in monomeric or multimeric form that are digested or otherwise produced from intact antibodies. In some embodiments of the invention, the Fc domain can be fused to FGF21 or FGF21 mutants (including truncated forms of FGF21 or FGF21 mutants), eg, via a covalent bond between the Fc domain and the FGF21 sequence. Such fusion proteins can form multimers via association of Fc domains, and both these fusion proteins and their multimers are an aspect of the present invention.

As used herein, the term "modified Fc fragment" shall mean an Fc fragment of an antibody comprising modified sequences. An Fc fragment is the portion of an antibody that includes portions of the CH2, CH3 and hinge regions. Modified Fc fragments can be derived, for example, from IgGl, IgG2, IgG3, or IgG4. FcLALA is a modified Fc fragment with LALA mutations (L234A, L235A) that triggers ADCC with lower efficiency and binds and activates human complement weakly. Hessell et al. 2007 Nature 449:101-104. Other modifications to Fc fragments are described, for example, in US Pat. No. 7,217,798, which is incorporated by reference for this purpose.

The term "acute myocardial infarction" refers to myocardial necrosis due to interruption of the blood supply to a part of the heart (eg, due to acute blockage of a coronary artery). The resulting ischemia and hypoxia can lead to damage or death (infarction) of myocardial tissue (myocardium) if left untreated for a prolonged period of time.

The term "atherosclerosis" is a vascular disease characterized by irregularly distributed lipid deposition in the intima of large and middle arteries, leading to narrowing of the arterial lumen and eventual progression to fibrosis and calcification. Lesions are usually focal and progress slowly and intermittently. Restriction of blood flow is responsible for most clinical manifestations, which vary with distribution and severity of lesions.

The term "cardiovascular disease" is a disease associated with the heart or blood vessels.

The term "cardiovascular risk" refers to the combination of factors associated with a higher probability of developing a cardiovascular event (eg, stroke or heart attack). Such factors may include, but are not limited to, weight, BMI, cholesterol levels, blood pressure, triglyceride levels, diet, exercise habits, age, gender, family history, obesity, diabetes, and/or other metabolic factors. Guidelines for assessing and managing cardiovascular risk have been described. For example, the American College of Cardiology (ACC) and the American Heart Association (AHA) have collaborated with the National Heart, Lung, and Blood Institute (NHLBI) and stakeholder and professional organizations to develop tools for assessing cardiovascular risk, lifestyle changes to reduce Cardiovascular risk, management of adult blood cholesterol, and clinical practice guidelines for managing adult overweight and obesity (Stone et al, 2014, Journal of the American College of Cardiology, 63(25, part B) 2889 -2934; DOI: 10.1016/j.jacc.2013.11.002, which is hereby incorporated by reference for this purpose). The World Health Organization also provides guidelines for assessing cardiovascular risk, eg, individuals with persistently elevated blood pressure ≥ 160/100 mm Hg, blood cholesterol ≥ 8 mmol/l, confirmed ischemic heart disease, or diabetes with concomitant renal disease.

The term "coronary heart disease" (also called coronary artery disease) is the narrowing of the small blood vessels that supply blood and oxygen to the heart.

The terms "diabetes" and "diabetic" refer to a progressive disorder of carbohydrate metabolism involving insufficient production or utilization of insulin, usually characterized by hyperglycemia and diabetes. The terms "pre-diabetic" and "pre-diabetic" refer to a state in which a subject does not have the features, symptoms, etc. typically observed in diabetes, but has features, symptoms, etc. that may progress to diabetes if untreated. The presence of these conditions can be determined using, for example, a fasting plasma glucose (FPG) test, an oral glucose tolerance test (OGTT), or a hemoglobin A1c (HbA1c) test. For both the FPG test and the OGTT, subjects are generally required to fast for at least 8 hours before starting the test. In the FPG test, the subject's blood glucose is measured after the fasting has ended; typically, the subject fasts overnight and the blood glucose is measured in the morning before the subject eats. Healthy subjects typically have FPG concentrations between about 90 and about 100 mg/dl, subjects with "pre-diabetes" typically have FPG concentrations between about 100 and about 125 mg/dl, and subjects with "diabetic" Subjects typically have FPG levels above about 126 mg/dl. In the OGTT, subjects' blood glucose was measured after fasting and again two hours after drinking a glucose-enriched beverage. Healthy subjects typically have blood glucose concentrations of less than about 140 mg/dl, prediabetic subjects typically have blood glucose concentrations of about 140 to about 199 mg/dl, and diabetic subjects two hours after consuming a glucose-enriched beverage Typically has a blood glucose concentration of about 200 mg/dl or higher. Although the above blood glucose values were related to human subjects, the proportions of normoglycemia, moderate hyperglycemia and marked hyperglycemia were different in mouse subjects. After a four-hour fast, healthy mouse subjects typically have FPG concentrations between about 100 mg/dl and about 150 mg/dl, and mouse subjects with "pre-diabetic" typically have between about 175 mg/dl and about 250 mg/dl FPG concentrations between dl, and mouse subjects with "diabetes" typically have FPG concentrations above about 250 mg/dl. For HbA1c testing, HbA1c levels <5.7% are generally considered to be within the normal range, HbA1c levels in the range of 5.7%-6.4% are generally considered pre-diabetic levels, and HbA1c levels of 6.5% or above are generally considered to be Diabetes level (see, eg, American Diabetes Association (ADA), Practice Guideline, Diabetes Care, 2018 Supplement 1, which is hereby incorporated by reference for this purpose).

The term "dyslipidemia" is a disorder of lipoprotein metabolism, including overproduction or deficiency of lipoproteins. Dyslipidemia may manifest as elevated total cholesterol, elevated low-density lipoprotein (LDL) cholesterol and triglyceride concentrations, and reduced blood high-density lipoprotein (HDL) cholesterol concentrations.

The term "glucose intolerance" or "impaired fasting glucose" (IFG) or "impaired glucose tolerance" (IGT) is a pre-diabetic state of dysglycemia associated with increased risk of cardiovascular pathology. The pre-diabetic condition prevents a subject from efficiently transferring glucose into cells and using it as an efficient fuel source, leading to elevated levels of glucose in the blood and some degree of insulin resistance.

The term "heart failure" (also known as congestive heart failure) is a condition in which the heart can no longer pump enough blood to the rest of the body.

The term "hyperglycemia" (also known as hyperglycemia) refers to a condition characterized by higher than normal amounts of glucose (a type of sugar) in the blood. Hyperglycemia can be diagnosed using methods known in the art, including measuring fasting blood glucose levels.

The term "hypertension or high blood pressure" refers to a condition in which systemic arterial blood pressure is temporarily or persistently elevated to levels that may induce cardiovascular damage or other adverse consequences. Hypertension is mandated to be defined as systolic blood pressure (SBP) above 140 mmHg or diastolic blood pressure (DBP) above 90 mmHg, but can also be defined according to published clinical guidelines. For example, the following blood pressure guidelines have been described: normal SBP < 120mmHg or DBP < 80mmHg; elevated SBP or DBP < 80mmHg in the range of 120-129mmHg; Hypertension (HTN) stage 1: SBP or DBP in the range of 130-139mmHg 80-89 mmHg; and HTN: SBP ≥ 140 or DBP ≥ 90.

The term "hypertriglyceridemia" refers to high blood levels of triglycerides (congestion). Elevated triglyceride levels are associated with atherosclerosis and predisposition to cardiovascular disease (eg, contributing to an increased risk of cardiovascular disease) even in the absence of hypercholesterolemia (high cholesterol levels). Typically, serum triglyceride levels in the range of 150 to 199 mg/dL [1.70 mmol/L to 2.25 mmol/L] are considered borderline high serum triglyceride levels; Serum triglyceride levels in the range of 5.64 mmol/L] are considered high serum triglyceride levels; and serum triglyceride levels in the range of 500 mg/dL [5.65 mmol/L] or higher are considered are very high triglyceride levels. In some aspects, these general guidelines may be adjusted based on the clinician's updated clinical guidelines.

The term "hypoglycemia" (also called hypoglycemia) occurs when blood sugar levels drop too low to provide enough energy for physical activity.

The term "hyperinsulinemia" is defined as higher than normal levels of insulin in the blood.

The term "insulin resistance" is defined as a state in which normal amounts of insulin produce a subnormal biological response.

The term "kidney disease" or nephropathy is any disease of the kidneys. Diabetic nephropathy refers to kidney damage caused by diabetes; in severe cases, it can lead to kidney failure, which is a major cause of morbidity and mortality in people with type 1 or type 2 diabetes.

The term "metabolic syndrome" refers to a combination of conditions (elevated blood pressure, high blood sugar, excess body fat around the waist, abnormal cholesterol or triglyceride levels) that increase the risk of heart disease, stroke and diabetes. For example, in most men, excess body fat around the waist was associated with a waist circumference of 40 inches or greater; high blood sugar was associated with post-fasting blood sugar/glucose levels of at least 110 milligrams per deciliter (mg/dl); high triglycerides Associated with levels in the bloodstream of at least 150 mg/dL; low HDL associated with levels below 40 mg/dL; and elevated blood pressure associated with levels of 130/85 mmHg or higher.

The term "myocardial infarction" (MI) (also known as a heart attack) is defined as the irreversible death (necrosis) of the heart muscle due to a prolonged lack of oxygen supply (ischemia).

The term "NAFLD" or "non-alcoholic fatty liver disease" is defined as a condition in which excess fat is stored in the liver. This fat accumulation is not caused by heavy alcohol use. When heavy alcohol use causes fat to build up in the liver, the condition is called alcoholic liver disease. Typically, there are two types of NAFLD: simple fatty liver and nonalcoholic steatohepatitis (NASH).

The term "NASH" or "non-alcoholic steatohepatitis" is defined as a form of non-alcoholic fatty liver disease (NAFLD) in which the patient has hepatitis - liver inflammation - liver cell damage in addition to liver fat. Inflammation and liver cell damage can lead to liver fibrosis or scarring. In some cases, NASH can lead to cirrhosis or liver cancer.

The term "simple fatty liver" (also known as nonalcoholic fatty liver (NAFL)) is a form of NAFLD in which a patient has liver fat, but little or no inflammation or liver cell damage. Simple fatty liver typically does not progress to cause liver damage or complications.

The term "pancreatitis" is inflammation of the pancreas.

"Obesity" is defined as abnormal or excessive fat accumulation that poses a risk to health. A rough population measure of obesity is body mass index (BMI=weight (kg)/[height (m)] ² ), which is a person's weight (in kilograms) divided by the square of his or her height (in meters). People with a BMI of 30 or more are generally considered obese. In general, people with a BMI of 25 or greater are considered overweight, for example, for Caucasians, but the guidelines can be adjusted for racial differences. For example, adjusted for ethnicity, a BMI ≥ 27.5 may be considered obese for Asian individuals (WHO Expert Consultation, 2004, Lancet, 363(9403):157-63). An Asian individual can be, for example, a subject of Asian descent or Asian ancestry.

The term "peripheral arterial disease" or "PAD" refers to a condition characterized by narrowing of peripheral arteries (arteries most commonly found in the legs) to the legs, stomach, arms and head. PAD is similar to coronary artery disease (CAD) in that both PAD and CAD are caused by atherosclerosis that narrows and blocks arteries in various critical areas of the body. The narrowing of the arteries is typically due to plaque, which can harden and narrow the arteries over time, restricting the flow of oxygen-rich blood to organs and other parts of the body.

The term "stroke" is any acute clinical event associated with impaired cerebral circulation lasting more than 24 hours, while shorter durations are considered transient ischemic attacks (TIAs). Stroke involves irreversible brain damage, and the type and severity of symptoms depend on the location and extent of circulating damaged brain tissue.

The term "Type 1 diabetes" or "T1DM" is a condition characterized by high blood sugar levels caused by a lack of insulin. The condition occurs when the body's immune system attacks and destroys insulin-producing beta cells in the pancreas. The pancreas then produces little or no insulin.

The term "type 2 diabetes" or "type 2 diabetes" or "T2D" or "T2DM" refers to a metabolic disease that causes sugar to build up in the bloodstream and is characterized by high blood sugar levels caused by a lack of insulin or the body's inability to use insulin effectively . Blood sugar levels are measured in milligrams per deciliter (mg/dL) or mmol/L.

The term "management or management or managing" is understood as the management and care of a patient for the purpose of combating a disease, condition or disorder, which does not result in a cure, but which results in one or more of the disease, condition or disorder Relief of symptoms and/or reduction in length of hospital stay.

The term "prevention or prevent or preventing" refers to the prophylactic administration to healthy subjects or subjects at risk of developing a disease, condition or disorder (eg, a subject considered to be pre-diabetic), to Prevent the development of one or more of the disorders mentioned herein. In addition, the term "prevention" can also include prophylactic administration to a patient in the pre-stage of the condition to be treated.

The term "treatment or treating or treating" is understood to mean the management and care of a patient for the purpose of combating a disease, condition or disorder, eg, reducing or ameliorating the progression, severity and/or duration of the disease, condition or disorder . In certain aspects, in the context of a disease, condition or disorder treatable by administration of FGF21 or FGF21 variant proteins provided herein, treatment as used herein can include, but is not limited to, one or more of the following: (i) reducing Triglyceride levels (blood/serum triglyceride levels), eg, to within manageable or normal ranges as determined by a clinician or clinical guidelines; (ii) weight loss, eg, to a level as determined by a clinician or Within the manageable range or normal range as determined by clinical guidelines, or at least about 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20 %, 25%, 30%, 35%, 40%, 50%, 55%, 60% or more; (iii) increase insulin-independent glucose uptake; (iv) decrease blood/serum glucose levels (eg, decrease To within manageable or normal range as determined by clinician or clinical guidelines, or decrease by at least about 5%, 10%, 20%, 30%, 40%, 45%, 50%, 55%, 60%, 70% %, 80%, 90%, or 95% or more); (v) improve lipoprotein profiles; (vi) increase HDL cholesterol (HDL-C) levels (eg, increase by at least about 5%, 10%, 20%) , 30%, 40%, 50%, 60%, 70%, 80%, 90% or more); (vii) reduce LDL cholesterol (LDL-C) and/or total cholesterol (total C), for example to Within the manageable or normal range as determined by a clinician or clinical guidelines, or a reduction of at least about 5%, 10%, 20%, 30%, 40%, or 50% or more; (viii) reduction in liver fat content , e.g., reduced to within a manageable or normal range as determined by a clinician or clinical guidelines, or reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more; and (ix ) to reduce the incidence of complications.

The term "therapeutically effective amount" or "effective amount" refers to the amount of a drug or therapeutic agent that will elicit the desired biological and/or medical response in a tissue, system or animal (including humans) being sought by a researcher or clinician. In the context of the FGF21 and FGF21 variant proteins provided herein, the desired biological and/or medical response may be one or more of the following: (i) reduction of triglyceride levels (blood/serum triglyceride levels) , for example, by at least about 5%, 10%, 20%, 30%, 40%, 45%, 50%, 55%, 60%, 70%, 80%, 90%, or 95% or more; ( ii) weight loss, eg, at least about 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30% %, 35%, 40%, 50%, 55%, 60% or more; (iii) increase insulin-independent glucose uptake; (iv) decrease blood/serum glucose levels (eg, decrease by at least about 5%, 10% %, 20%, 30%, 40%, 45%, 50%, 55%, 60%, 70%, 80%, 90%, or 95% or more); (v) improve lipoprotein profiles; (vi) ) increase HDL cholesterol (HDL-C) levels (eg, by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more); (vii) reducing LDL cholesterol (LDL-C) and/or total cholesterol (total C), eg, by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more; and (viii) ) reduces liver fat content, eg, by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more. In certain embodiments, triglyceride levels can be reduced by at least about 40% or at least about 50%. In certain embodiments, triglyceride levels can be reduced by at least about 40%. In certain embodiments, triglyceride levels can be reduced by at least about 50%. In some embodiments, liver fat content can be reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more. In some embodiments, liver fat content can be reduced by at least about 5%. In some embodiments, liver fat content can be reduced by at least about 10%. In some embodiments, liver fat content can be reduced by at least about 20%. In some embodiments, liver fat content can be reduced by at least about 30%. In some embodiments, liver fat content can be reduced by at least about 40%. In some embodiments, liver fat content can be reduced by at least about 50%. In some embodiments, liver fat content can be reduced to within a manageable or normal range as determined by a clinician or clinical guidelines.

As used herein, the singular forms "a/an" and "the" include plural referents unless the content clearly dictates otherwise. Thus, for example, reference to an "antibody" includes a mixture of two or more such antibodies.

As used herein, the term "about" or "approximately" means +/- 20%, more preferably +/- 10%, or still more preferably +/- 5% of a value.

The terms "polypeptide" and "protein" are used interchangeably and refer to a polymeric form of amino acids of any length, which may include encoded and non-encoded amino acids, naturally occurring and non-naturally occurring amino acids, chemically or biochemically modified or Derivatized amino acids, and polypeptides with modified peptide backbones. The term includes fusion proteins including, but not limited to, fusion proteins with heterologous amino acid sequences, fusions with heterologous and homologous leader sequences (with or without N-terminal methionine residues); immunolabeled protein; etc.

The term "fragment" as used herein refers to a physically contiguous portion of the primary structure of a biomolecule. In the case of a protein, a portion is defined by a contiguous portion of the amino acid sequence of the protein, and refers to at least 3-5 amino acids, at least 8-10 amino acids, at least 11-15 amino acids, at least 17-24 amino acids, at least 25-30 amino acids, and at least 30-45 amino acids. In the case of an oligonucleotide, a portion is defined by a contiguous portion of the nucleic acid sequence of the oligonucleotide, and refers to at least 9-15 nucleotides, at least 18-30 nucleotides, at least 33-45 nucleotides nucleotides, at least 48-72 nucleotides, at least 75-90 nucleotides, and at least 90-130 nucleotides. In some embodiments, the portion of the biomolecule is biologically active. In the context of FGF21 polypeptides, FGF21 polypeptide fragments do not comprise the entire FGF21 polypeptide sequence set forth in SEQ ID NO:3.

The terms "individual", "subject", "host" and "patient" are used interchangeably and refer to any subject, particularly a human, in need of diagnosis, treatment or therapy. Other subjects may include cows, dogs, cats, guinea pigs, rabbits, rats, mice, horses, and the like. In some preferred embodiments, the subject is a human.

As used herein, the term "sample" refers to biological material from a patient. The samples assayed by the present invention are not limited to any particular type. As non-limiting examples, samples include single cells, multiple cells, tissues, tumors, biological fluids, biomolecules, or supernatants or extracts of any of the foregoing. Examples include tissue removed for biopsy, tissue removed during resection, blood, urine, lymphoid tissue, lymph, cerebrospinal fluid, mucus, and stool samples. The sample used will vary depending on the assay format, the detection method and the nature of the tumor, tissue, cell or extract to be assayed. Methods for preparing samples are well known in the art and can be easily adapted to obtain samples compatible with the method used.

The term "pharmaceutically acceptable carrier" refers to a carrier used to administer therapeutic agents such as antibodies or polypeptides, genes and other therapeutic agents. The term refers to any pharmaceutical carrier that does not by itself induce the production of antibodies detrimental to the individual receiving the composition, and which can be administered without undue toxicity. Suitable carriers may be large, slowly metabolized macromolecules such as proteins, polysaccharides, polylactic acid, polyglycolic acid, polymeric amino acids, amino acid copolymers, lipid aggregates and/or inactive viral particles. Such vectors are well known to those of ordinary skill in the art. Pharmaceutically acceptable carriers in therapeutic compositions can include liquids such as water, saline, glycerol and/or ethanol. Auxiliary substances such as wetting or emulsifying agents, pH buffering substances and the like may also be present in such vehicles.

treatment method

Provided herein are methods and pharmaceutical compositions for the treatment, prevention, or management of metabolic or cardiovascular disorders (eg, alleviating one or more symptoms thereof), including at doses in the range of 100 mg to 600 mg, eg, once daily, every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month) to subjects in need thereof to administer human FGF21 protein Variants (eg, Fc-FGF21 mutant fusion proteins, such as V103 (SEQ ID NO: 11)). In particular aspects, the human FGF21 protein variant (eg, an Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)) is administered at a dose ranging from 200 mg to 400 mg. In particular aspects, the human FGF21 protein variant (eg, an Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)) is administered at a dose of about 200 mg, 250 mg, 300 mg, 350 mg, or 400 mg, eg, once a week, Administration is every 2 weeks, every 3 weeks, or every 4 weeks (or monthly). In particular aspects, the human FGF21 protein variant (eg, an Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)) is administered every 4 weeks (or monthly). In certain aspects, the human FGF21 protein variant (eg, V103 (SEQ ID NO: 11)) is administered every 3 weeks, every 2 weeks, or once a week.

In one particular aspect, provided herein are methods of treating, preventing, or managing (eg, alleviating one or more symptoms of) a metabolic disorder or a cardiovascular disorder (eg, obesity or hypertriglyceridemia), the method comprising A human FGF21 protein variant (e.g., an Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)). In one particular aspect, provided herein are methods of treating, preventing, or managing (eg, alleviating one or more symptoms of) a metabolic disorder or a cardiovascular disorder (eg, obesity or hypertriglyceridemia), the method comprising Administering a human FGF21 protein variant (eg, an Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)).

In one particular aspect, provided herein are methods of treating, preventing, or managing (eg, alleviating one or more symptoms of) a metabolic disorder or a cardiovascular disorder (eg, obesity or hypertriglyceridemia), the method comprising A human FGF21 protein variant (e.g., an Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)). In one particular aspect, provided herein are methods of treating, preventing, or managing (eg, alleviating one or more symptoms of) a metabolic disorder or a cardiovascular disorder (eg, obesity or hypertriglyceridemia), the method comprising Administering a human FGF21 protein variant (eg, an Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)).

In one particular aspect, provided herein are methods of treating, preventing, or managing (eg, alleviating one or more symptoms of) a metabolic disorder or a cardiovascular disorder (eg, obesity or hypertriglyceridemia), the method comprising A human FGF21 protein variant (e.g., an Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)). In one particular aspect, provided herein are methods of treating, preventing, or managing (eg, alleviating one or more symptoms of) a metabolic disorder or a cardiovascular disorder (eg, obesity or hypertriglyceridemia), the method comprising Administering a human FGF21 protein variant (eg, an Fc-FGF21 mutant fusion protein, such as V103 (SEQ ID NO: 11)).

In one particular aspect, provided herein are methods of treating, preventing, or managing (eg, alleviating one or more symptoms of) a metabolic disorder or a cardiovascular disorder (eg, obesity or hypertriglyceridemia), the method comprising A human FGF21 protein variant (eg, an Fc-FGF21 mutant fusion protein) is administered to a subject in need thereof at a dose of at least about 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, or 350 mg once a week or once every two weeks , such as V103 (SEQ ID NO: 11)). In one particular aspect, provided herein are methods of treating, preventing, or managing (eg, alleviating one or more symptoms of) a metabolic disorder or a cardiovascular disorder (eg, obesity or hypertriglyceridemia), the method comprising A human FGF21 protein variant (e.g., an Fc IgG) is administered to a subject in need thereof at a dose of at least about 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, or 350 mg every three weeks or every four weeks (or monthly). -FGF21 mutant fusion proteins such as V103 (SEQ ID NO: 11)).

In one aspect, non-limiting examples of metabolic or cardiovascular disorders to be treated, prevented or managed by the methods provided herein include obesity, hypertriglyceridemia, dyslipidemia, non-alcoholic fatty liver disease (NAFLD) , nonalcoholic steatohepatitis (NASH), diabetes (eg, type 2 diabetes (T2DM) and type 1 diabetes (T1DM)), diseases associated with severely inactivating mutations in the insulin receptor, pancreatitis, insulin resistance, high Insulinemia, glucose intolerance, hyperglycemia, metabolic syndrome, hypertension, cardiovascular disease, acute myocardial infarction, atherosclerosis, peripheral arterial disease, stroke, heart failure, coronary heart disease, kidney disease, diabetes complications disease, neuropathy, and gastroparesis.

In one aspect, non-limiting examples of metabolic or cardiovascular disorders to be treated, prevented or managed by the methods provided herein include obesity, hypertriglyceridemia and heart disease risk, insulin resistance (eg, having insulin receptors) patients with genetic mutations) and lipodystrophy, diabetes (eg, T1DM or T2DM), and nonalcoholic fatty liver disease (NAFLD)/nonalcoholic steatohepatitis (NASH).

In one aspect, provided herein is a method of treating, preventing or managing a metabolic disorder (eg, obesity) or a cardiovascular disorder, the method comprising at a dose in the range of 100 mg to 600 mg (or more specifically, at a dose of 100 mg to 300 mg or Doses in the range of 200 mg to 400 mg) such as once daily, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month) ) to administer (eg, subcutaneously (sc)) a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) to a subject in need thereof. In one aspect, provided herein is a method of treating, preventing, or managing a metabolic disorder (eg, obesity) or a cardiovascular disorder, the method comprising at about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, Doses of 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, e.g. once daily, every 2, 3, 4, 5 or 6 A human FGF21 protein variant (e.g., Fc fusion proteins such as V103 (SEQ ID NO: 11)). In one particular aspect, provided herein is a method of treating, preventing or managing a metabolic disorder (eg, obesity) or a cardiovascular disorder, the method comprising administering to a subject in need thereof a dose of about 300 mg every 4 weeks (monthly). The subject is administered (eg, sc) a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) subcutaneously. In one particular aspect, provided herein is a method of treating, preventing or managing a metabolic disorder (eg, obesity) or a cardiovascular disorder, the method comprising subcutaneously administering to a subject in need thereof a dose of about 300 mg every 3 weeks (eg sc) human FGF21 protein variants (eg, Fc fusion proteins such as V103 (SEQ ID NO: 11)). In one particular aspect, provided herein is a method of treating, preventing or managing a metabolic disorder (eg, obesity) or a cardiovascular disorder, the method comprising subcutaneously administering to a subject in need thereof a dose of about 300 mg every 2 weeks (eg sc) human FGF21 protein variants (eg, Fc fusion proteins such as V103 (SEQ ID NO: 11)). In one particular aspect, provided herein is a method of treating, preventing, or managing a metabolic disorder (eg, obesity) or a cardiovascular disorder, the method comprising administering to a dose in a range of about 200 mg to 250 mg every 3 weeks in need thereof The subject is administered a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) subcutaneously. In particular aspects, the methods provided herein are for treating human subjects with hypercholesterolemia (eg, primary hypercholesterolemia) or dyslipidemia (eg, mixed dyslipidemia). In certain aspects, the methods provided herein are for treating human subjects suffering from hypertriglyceridemia, particularly severe hypertriglyceridemia. In certain aspects, the methods provided herein are for treating a human subject with type 2 diabetes or an obese human subject. In particular aspects, the methods provided herein are for treating a human subject between the ages of 18 and 55 who, prior to administration of the human FGF21 protein variant, has (i) in the range of 30-45 kg/m ⁽ BMI (with race-adjusted greater than or equal to 27.5 kg/m ² BMI for Asian subjects), inclusive, and (ii) glycerol in the range of 150 to 500 mg/dL (1.69-5.65 mmol/L) Triester levels.

In particular aspects, provided herein are methods of treating, preventing, or managing hypercholesterolemia, dyslipidemia (eg, mixed dyslipidemia), or hypertriglyceridemia (eg, alleviating one or more symptoms thereof), the methods Included in doses in the range of 100 mg to 600 mg (or more specifically, in the range of 100 mg to 300 mg or 200 mg to 400 mg) such as once a day, once every 2, 3, 4, 5 or 6 days, once a week , once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month) to a subject in need thereof to administer a human FGF21 protein variant (e.g., an Fc fusion protein such as V103 (SEQ ID NO: 11)). In one particular aspect, provided herein are methods of treating, preventing, or managing hypercholesterolemia, dyslipidemia (eg, mixed dyslipidemia), or hypertriglyceridemia (eg, alleviating one or more symptoms thereof), which The method includes at about Doses of 330 mg, 340 mg, or 350 mg such as once daily, once every 2, 3, 4, 5 or 6 days, once weekly, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month) A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered to a subject in need thereof. In a certain aspect, provided herein are methods of treating, preventing, or managing hypercholesterolemia, dyslipidemia (eg, mixed dyslipidemia), or hypertriglyceridemia (eg, alleviating one or more symptoms thereof), the The method comprises subcutaneously administering a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg once every 4 weeks (or once a month) . In a certain aspect, provided herein are methods of treating, preventing, or managing hypercholesterolemia, dyslipidemia (eg, mixed dyslipidemia), or hypertriglyceridemia (eg, alleviating one or more symptoms thereof), the The method comprises subcutaneously administering a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg once every 3 weeks. In a certain aspect, provided herein are methods of treating, preventing, or managing hypercholesterolemia, dyslipidemia (eg, mixed dyslipidemia), or hypertriglyceridemia (eg, alleviating one or more symptoms thereof), the The method comprises subcutaneously administering a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg once every 2 weeks. In a certain aspect, provided herein are methods of treating, preventing, or managing hypercholesterolemia, dyslipidemia (eg, mixed dyslipidemia), or hypertriglyceridemia (eg, alleviating one or more symptoms thereof), the The method comprises subcutaneously administering a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg once per week. In a certain aspect, provided herein are methods of treating, preventing, or managing hypercholesterolemia, dyslipidemia (eg, mixed dyslipidemia), or hypertriglyceridemia (eg, alleviating one or more symptoms thereof), the The method comprises subcutaneously administering a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO) to a subject in need thereof at a dose in the range of about 200 mg to 250 mg once every 3 weeks or once every 2 weeks. :11)). In particular aspects, the methods provided herein are for treating human subjects with hypercholesterolemia (eg, primary hypercholesterolemia) or dyslipidemia (eg, mixed dyslipidemia). In certain aspects, the methods provided herein are for treating human subjects suffering from hypertriglyceridemia, particularly severe hypertriglyceridemia. In certain aspects, the methods provided herein are for treating a human subject with type 2 diabetes or an obese human subject. In particular aspects, the methods provided herein are for treating a human subject between the ages of 18 and 55 who, prior to administration of the human FGF21 protein variant, has (i) in the range of 30-45 kg/m ⁽ BMI (with race-adjusted greater than or equal to 27.5 kg/m ² BMI for Asian subjects), inclusive, and (ii) glycerol in the range of 150 to 500 mg/dL (1.69-5.65 mmol/L) Triester levels.

In particular aspects, provided herein are methods of reducing body weight (eg, reducing body weight by at least 5%, 6%, 7%, 8%, 9%, 10%, 15%, or 20% or more) comprising at a dose in the range of 100 mg to 600 mg (or more specifically, in a dose in the range of 200 mg to 400 mg or 100 mg to 300 mg) such as once a day, once every 2, 3, 4, 5 or 6 days, once a week, A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)). In a certain aspect, provided herein is a method of reducing body weight (eg, reducing body weight by at least 5%, 6%, 7%, 8%, 9%, 10%, 15%, or 20% or more), the method Including at about 100mg, 110mg, 120mg, 130mg, 140mg, 150mg, 160mg, 170mg, 180mg, 190mg, 200mg, 210mg, 220mg, 230mg, 240mg, 250mg, 260mg, 270mg, 280mg, 290mg, 300mg, 310mg, 320mg, 330mg , 340 mg, or 350 mg doses such as once daily, once every 2, 3, 4, 5, or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month) to A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered to a subject in need thereof. In a certain aspect, provided herein is a method of reducing body weight (eg, reducing body weight by at least 5%, 6%, 7%, 8%, 9%, 10%, 15%, or 20% or more), the method This includes subcutaneous administration of a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of approximately 300 mg every 4 weeks (or monthly). In a certain aspect, provided herein is a method of reducing body weight (eg, reducing body weight by at least 5%, 6%, 7%, 8%, 9%, 10%, 15%, or 20% or more), the method This includes subcutaneous administration of a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of approximately 300 mg once every 3 weeks. In a certain aspect, provided herein is a method of reducing body weight (eg, reducing body weight by at least 5%, 6%, 7%, 8%, 9%, 10%, 15%, or 20% or more), the method This includes subcutaneous administration of a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of approximately 300 mg once every 2 weeks. In a certain aspect, provided herein is a method of reducing body weight (eg, reducing body weight by at least 5%, 6%, 7%, 8%, 9%, 10%, 15%, or 20% or more), the method This includes subcutaneous administration of a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg once a week. In a certain aspect, provided herein is a method of reducing body weight (eg, reducing body weight by at least 5%, 6%, 7%, 8%, 9%, 10%, 15%, or 20% or more), the method This includes subcutaneous administration of a human FGF21 protein variant (e.g., an Fc fusion protein such as V103 (SEQ ID NO: 11)). In particular aspects, the methods provided herein are for treating human subjects with hypercholesterolemia (eg, primary hypercholesterolemia) or dyslipidemia (eg, mixed dyslipidemia). In certain aspects, the methods provided herein are for treating human subjects suffering from hypertriglyceridemia, particularly severe hypertriglyceridemia. In certain aspects, the methods provided herein are for treating a human subject with type 2 diabetes or an obese human subject. In certain aspects, the methods provided herein are for the treatment of a human subject between 18 and 55 years of age who, when screened prior to administration of a human FGF21 protein variant, has (i ⁾ a 30-45 kg/m BMI within the range (inclusive) (with race-adjusted greater than or equal to 27.5 kg/ ^m2 BMI for Asian subjects), and (ii) within the range of 150 to 500 mg/dL (1.69-5.65 mmol/L) internal triglyceride levels.

In particular aspects, provided herein are methods of treating, preventing, or managing obesity (eg, alleviating one or more symptoms thereof), the methods comprising at a dose ranging from 100 mg to 600 mg (or more specifically, at a dose ranging from 200 mg to 600 mg) 400 mg or doses in the range of 100 mg to 300 mg) such as once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or every monthly) to a subject in need thereof a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered. In one particular aspect, provided herein is a method of treating, preventing, or managing obesity (eg, alleviating one or more symptoms thereof), the method comprising administering at about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg , 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg at doses such as once daily every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month) to a subject in need thereof administering a human FGF21 protein variant (e.g., Fc Fusion proteins such as V103 (SEQ ID NO: 11)). In a certain aspect, provided herein is a method of treating, preventing, or managing obesity (eg, alleviating one or more symptoms thereof), the method comprising administering to it at a dose of about 300 mg every 4 weeks (once a month) A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered subcutaneously to a subject in need thereof. In a certain aspect, provided herein is a method of treating, preventing or managing obesity (eg, alleviating one or more symptoms thereof), the method comprising administering to a subject in need thereof a dose of about 300 mg once every 3 weeks A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered subcutaneously. In a certain aspect, provided herein is a method of treating, preventing or managing obesity (eg, alleviating one or more symptoms thereof), the method comprising administering to a subject in need thereof a dose of about 300 mg once every 2 weeks A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered subcutaneously. In a certain aspect, provided herein is a method of treating, preventing, or managing obesity (eg, alleviating one or more symptoms thereof), the method comprising administering to a subject in need thereof a dose of about 300 mg once every 1 week A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered subcutaneously. In a certain aspect, provided herein is a method of treating, preventing or managing obesity (eg, alleviating one or more symptoms thereof), the method comprising once every 3 weeks or every 2 at a dose in the range of about 200 mg to 250 mg A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered subcutaneously once a week to a subject in need thereof. In particular aspects, the methods provided herein are for treating human subjects with hypercholesterolemia (eg, primary hypercholesterolemia) or dyslipidemia (eg, mixed dyslipidemia). In certain aspects, the methods provided herein are for treating human subjects suffering from hypertriglyceridemia, particularly severe hypertriglyceridemia. In certain aspects, the methods provided herein are for treating a human subject with type 2 diabetes or an obese human subject. In certain aspects, the methods provided herein are for the treatment of a human subject between 18 and 55 years of age who, when screened prior to administration of a human FGF21 protein variant, has (i ⁾ a 30-45 kg/m BMI within the range (inclusive) (with race-adjusted greater than or equal to 27.5 kg/ ^m2 BMI for Asian subjects), and (ii) within the range of 150 to 500 mg/dL (1.69-5.65 mmol/L) internal triglyceride levels.

In particular aspects, provided herein are methods of treating, preventing, or managing NASH (eg, alleviating one or more symptoms thereof) comprising at a dose in the range of 100 mg to 600 mg (or more specifically, at a dose of 200 mg to 400 mg) or doses in the range of 100 mg to 300 mg) such as once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or monthly once) a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered to a subject in need thereof. In one particular aspect, provided herein is a method of treating, preventing, or managing NASH (eg, alleviating one or more symptoms thereof), the method comprising at about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, Doses of 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, e.g. once daily, every 2, 3, 4, 5 or once every 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month) to a subject in need thereof to administer a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)). In a certain aspect, provided herein is a method of treating, preventing or managing NASH (eg, alleviating one or more symptoms thereof), the method comprising administering to it at a dose of about 300 mg every 4 weeks (or once a month) A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered subcutaneously to a subject in need thereof. In a certain aspect, provided herein is a method of treating, preventing or managing NASH (eg, alleviating one or more symptoms thereof) comprising administering to a subject in need thereof a dose of about 300 mg once every 3 weeks Human FGF21 protein variants (eg, Fc fusion proteins such as V103 (SEQ ID NO: 11)) are administered subcutaneously. In a certain aspect, provided herein is a method of treating, preventing or managing NASH (eg, alleviating one or more symptoms thereof) comprising administering to a subject in need thereof a dose of about 300 mg once every 2 weeks Human FGF21 protein variants (eg, Fc fusion proteins such as V103 (SEQ ID NO: 11)) are administered subcutaneously. In a certain aspect, provided herein is a method of treating, preventing or managing NASH (eg, alleviating one or more symptoms thereof) comprising subcutaneously subcutaneously administering a dose of about 300 mg once a week to a subject in need thereof A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered. In a certain aspect, provided herein is a method of treating, preventing or managing NASH (eg, alleviating one or more symptoms thereof) comprising once every 3 weeks or every 2 weeks at a dose in the range of about 200 mg to 250 mg A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered subcutaneously at one time to a subject in need thereof. In particular aspects, the methods provided herein are for treating human subjects with hypercholesterolemia (eg, primary hypercholesterolemia) or dyslipidemia (eg, mixed dyslipidemia). In certain aspects, the methods provided herein are for treating human subjects suffering from hypertriglyceridemia, particularly severe hypertriglyceridemia. In certain aspects, the methods provided herein are for treating a human subject with type 2 diabetes or an obese human subject. In particular aspects, the methods provided herein are for treating a human subject between the ages of 18 and 55 who, prior to administration of the human FGF21 protein variant, has (i) in the range of 30-45 kg/m ⁽ BMI (with race-adjusted greater than or equal to 27.5 kg/m ² BMI for Asian subjects), inclusive, and (ii) glycerol in the range of 150 to 500 mg/dL (1.69-5.65 mmol/L) Triester levels.

In particular aspects, provided herein are methods of treating, preventing, or managing NAFLD (eg, alleviating one or more symptoms thereof) comprising at a dose in the range of 100 mg to 600 mg (or more specifically, at a dose of 200 mg to 400 mg) or doses in the range of 100 mg to 300 mg) such as once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or monthly once) a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered to a subject in need thereof. In one particular aspect, provided herein is a method of treating, preventing, or managing NAFLD (eg, alleviating one or more symptoms thereof), the method comprising administering at about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, Doses of 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg, e.g. once daily, every 2, 3, 4, 5 or once every 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month) to a subject in need thereof to administer a human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)). In a certain aspect, provided herein is a method of treating, preventing, or managing NAFLD (eg, alleviating one or more symptoms thereof), the method comprising administering to the NAFLD once every 4 weeks (or once a month) at a dose of about 300 mg A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered subcutaneously to a subject in need thereof. In a certain aspect, provided herein is a method of treating, preventing or managing NAFLD (eg, alleviating one or more symptoms thereof) comprising administering to a subject in need thereof a dose of about 300 mg once every 3 weeks Human FGF21 protein variants (eg, Fc fusion proteins such as V103 (SEQ ID NO: 11)) are administered subcutaneously. In a certain aspect, provided herein is a method of treating, preventing or managing NAFLD (eg, alleviating one or more symptoms thereof), the method comprising administering to a subject in need thereof a dose of about 300 mg once every 2 weeks Human FGF21 protein variants (eg, Fc fusion proteins such as V103 (SEQ ID NO: 11)) are administered subcutaneously. In a certain aspect, provided herein is a method of treating, preventing, or managing NAFLD (eg, alleviating one or more symptoms thereof) comprising subcutaneously subcutaneously administering to a subject in need thereof a dose of about 300 mg once weekly A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered. In a certain aspect, provided herein is a method of treating, preventing or managing NAFLD (eg, alleviating one or more symptoms thereof) comprising once every 3 weeks or every 2 weeks at a dose in the range of about 200 mg to 250 mg A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered subcutaneously at one time to a subject in need thereof. In particular aspects, the methods provided herein are for treating human subjects with hypercholesterolemia (eg, primary hypercholesterolemia) or dyslipidemia (eg, mixed dyslipidemia). In certain aspects, the methods provided herein are for treating human subjects suffering from hypertriglyceridemia, particularly severe hypertriglyceridemia. In certain aspects, the methods provided herein are for treating a human subject with type 2 diabetes or an obese human subject. In particular aspects, the methods provided herein are for treating a human subject between the ages of 18 and 55 who, prior to administration of the human FGF21 protein variant, has (i) in the range of 30-45 kg/m ⁽ BMI (with race-adjusted greater than or equal to 27.5 kg/m ² BMI for Asian subjects), inclusive, and (ii) glycerol in the range of 150 to 500 mg/dL (1.69-5.65 mmol/L) Triester levels.

In particular aspects, provided herein are methods of reducing fat or lipids in the liver, the methods comprising at a dose in the range of 100 mg to 600 mg (or more specifically, at a dose in the range of 200 mg to 400 mg or 100 mg to 300 mg), eg Daily, every 2, 3, 4, 5, or 6 days, weekly, every 2 weeks, every 3 weeks, or every 4 weeks (or monthly) to subjects in need (eg, a subject with NASH or NAFLD) is administered a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)). In a particular aspect, provided herein is a method of reducing fat or lipid in the liver, the method comprising administering at about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg , 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg at doses such as once a day, every 2, 3, 4, 5 or 6 days, once a week, every 2 A human FGF21 protein variant (e.g., Fc fusion proteins such as V103 (SEQ ID NO: 11)). In a certain aspect, provided herein is a method of reducing fat or lipids in the liver, the method comprising administering to a subject in need thereof (eg, suffering from subjects with NASH or NAFLD) administered a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) subcutaneously. In a certain aspect, provided herein is a method of reducing fat or lipids in the liver, the method comprising administering to a subject in need thereof (eg, a subject suffering from NASH or NAFLD) at a dose of about 300 mg once every 3 weeks or) subcutaneously administer a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)). In a certain aspect, provided herein is a method of reducing fat or lipids in the liver, the method comprising administering to a subject in need thereof (eg, a subject suffering from NASH or NAFLD) at a dose of about 300 mg once every 2 weeks or) subcutaneously administer a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)). In a certain aspect, provided herein is a method of reducing fat or lipids in the liver, the method comprising administering to a subject in need thereof (eg, a subject suffering from NASH or NAFLD) once a week at a dose of about 300 mg ) subcutaneously administer a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)). In a certain aspect, provided herein is a method of reducing fat or lipids in the liver, the method comprising administering to a subject in need thereof ( For example, a subject with NASH or NAFLD) administers a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) subcutaneously. In certain aspects, the methods provided herein are for treating human subjects with NASH or NAFLD. In particular aspects, the methods provided herein are for treating human subjects with hypercholesterolemia (eg, primary hypercholesterolemia) or dyslipidemia (eg, mixed dyslipidemia). In certain aspects, the methods provided herein are for treating human subjects suffering from hypertriglyceridemia, particularly severe hypertriglyceridemia. In certain aspects, the methods provided herein are for treating a human subject with type 2 diabetes or an obese human subject. In certain aspects, the methods provided herein are for the treatment of a human subject between 18 and 55 years of age who, when screened prior to administration of a human FGF21 protein variant, has (i ⁾ a 30-45 kg/m BMI within the range (inclusive) (with race-adjusted greater than or equal to 27.5 kg/ ^m2 BMI for Asian subjects), and (ii) within the range of 150 to 500 mg/dL (1.69-5.65 mmol/L) internal triglyceride levels.

In particular aspects, provided herein are methods of lowering elevated LDL cholesterol (LDL-C), total cholesterol (total C), triglycerides, and/or Apo B, the methods comprising at doses ranging from 100 mg to 600 mg, eg Daily, every 2, 3, 4, 5, or 6 days, weekly, every 2 weeks, every 3 weeks, or every 4 weeks (or monthly) to subjects in need A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered. In particular aspects, provided herein are methods of lowering elevated LDL-C, total C, triglycerides, and/or Apo B, the methods comprising at a dose in the range of 100 mg to 600 mg (or more specifically, at a dose ranging from 200 mg to 600 mg) 400 mg or doses ranging from 100 mg to 300 mg) administer a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) to a subject in need thereof. In a specific aspect, provided herein is a method of reducing elevated LDL-C, total C, triglycerides and/or Apo B, the method comprising at about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg , 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg at doses such as once daily every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks (or once a month) to a subject in need thereof administering a human FGF21 protein variant (e.g., Fc Fusion proteins such as V103 (SEQ ID NO: 11)). In a certain aspect, provided herein is a method of lowering elevated LDL-C, total C, triglycerides, and/or Apo B, the method comprising administering to the drug at a dose of about 300 mg every 4 weeks (once a month) A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered subcutaneously to a subject in need thereof. In a certain aspect, provided herein is a method of reducing elevated LDL-C, total C, triglycerides and/or Apo B, the method comprising administering to a subject in need thereof a dose of about 300 mg once every 3 weeks A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered subcutaneously. In a certain aspect, provided herein is a method of reducing elevated LDL-C, total C, triglycerides and/or Apo B, the method comprising administering to a subject in need thereof a dose of about 300 mg once every 2 weeks A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered subcutaneously. In a certain aspect, provided herein is a method of reducing elevated LDL-C, total C, triglycerides, and/or Apo B, the method comprising administering to a subject in need thereof a dose of about 300 mg once weekly Human FGF21 protein variants (eg, Fc fusion proteins such as V103 (SEQ ID NO: 11)) are administered subcutaneously. In a certain aspect, provided herein is a method of lowering elevated LDL-C, total C, triglycerides, and/or Apo B, the method comprising once every 3 weeks or every 2 at a dose in the range of about 200 mg to 250 mg A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered subcutaneously once a week to a subject in need thereof. In a specific aspect, the methods provided herein reduce elevated LDL-C, total C, triglycerides and/or Apo B by at least about 5%, 6%, 7%, 8%, 9%, 10%, 11% %, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, or 90% or more. In particular aspects, the methods provided herein are for treating human subjects with hypercholesterolemia (eg, primary hypercholesterolemia) or dyslipidemia (eg, mixed dyslipidemia). In certain aspects, the methods provided herein are for treating human subjects suffering from hypertriglyceridemia, particularly severe hypertriglyceridemia. In certain aspects, the methods provided herein are for treating a human subject with type 2 diabetes or an obese human subject. In certain aspects, the methods provided herein are for the treatment of a human subject between 18 and 55 years of age who, when screened prior to administration of a human FGF21 protein variant, has (i ⁾ a 30-45 kg/m BMI within the range (inclusive) (with race-adjusted greater than or equal to 27.5 kg/ ^m2 BMI for Asian subjects), and (ii) within the range of 150 to 500 mg/dL (1.69-5.65 mmol/L) internal triglyceride levels.

In certain aspects, provided herein is increasing HDL-C (eg, increasing HDL-C by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% ) method comprising at a dose in the range of 100 mg to 600 mg, for example, once daily, once every 2, 3, 4, 5 or 6 days, once weekly, once every 2 weeks, once every 3 weeks, or once every 4 weeks A therapeutically effective amount of a fusion protein comprising a human FGF21 protein variant (eg, an Fc fusion protein, such as V103 (SEQ ID NO: 11)) is administered weekly (or monthly) to a subject in need thereof. In certain aspects, provided herein is increasing HDL-C (eg, increasing HDL-C by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% ), the method comprising administering to a subject in need thereof a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) at a dose ranging from about 200 mg to 400 mg. In one particular aspect, provided herein is increasing HDL-C (eg, increasing HDL-C by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%) %) method, the method comprising taking about Doses of 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg such as once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered (or once a month) to a subject in need thereof. In a certain aspect, provided herein is increasing HDL-C (eg, increasing HDL-C by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%) %) method comprising subcutaneously administering to a subject in need thereof a human FGF21 protein variant ( For example, Fc fusion proteins such as V103 (SEQ ID NO: 11)). In a certain aspect, provided herein is increasing HDL-C (eg, increasing HDL-C by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%) %), the method comprising subcutaneously administering to a subject in need thereof a human FGF21 protein variant (e.g., an Fc fusion protein, Fc fusion protein, Such as V103 (SEQ ID NO: 11)). In particular aspects, the methods provided herein are for treating human subjects with hypercholesterolemia (eg, primary hypercholesterolemia) or dyslipidemia (eg, mixed dyslipidemia). In certain aspects, the methods provided herein are for treating human subjects suffering from hypertriglyceridemia, particularly severe hypertriglyceridemia. In certain aspects, the methods provided herein are for treating a human subject with type 2 diabetes or an obese human subject. In certain aspects, the methods provided herein are for the treatment of a human subject between 18 and 55 years of age who, when screened prior to administration of a human FGF21 protein variant, has (i ⁾ a 30-45 kg/m BMI within the range (inclusive) (with race-adjusted greater than or equal to 27.5 kg/ ^m2 BMI for Asian subjects), and (ii) within the range of 150 to 500 mg/dL (1.69-5.65 mmol/L) internal triglyceride levels.

In certain aspects, provided herein is reducing triglyceride levels (eg, fasting triglyceride levels) (eg, reducing blood/serum triglyceride levels by at least 20%, 30%, 40%, 50%, 60%, 70%) , 80%, or 90% or more), the method comprising at a dose in the range of 100 mg to 600 mg, for example, once daily, once every 2, 3, 4, 5 or 6 days, once a week, every 2 weeks A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered to a subject in need thereof once, once every 3 weeks, or once every 4 weeks (or once a month) . In certain aspects, provided herein is reducing triglyceride levels (eg, reducing blood/serum triglyceride levels by at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% or more), the method comprising administering to a subject in need thereof a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11) at a dose in a range of about 200 mg to 400 mg) ). In a particular aspect, provided herein is reducing triglyceride levels (eg, fasting triglyceride levels) (eg, reducing blood/serum triglyceride levels by at least 20%, 30%, 40%, 50%, 60%, 70%) %, 80%, or 90% or more), the method comprising at about Doses of 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg e.g. once daily, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered to a subject in need thereof once every 3 weeks, or once every 4 weeks (or monthly). In a certain aspect, provided herein is reducing triglyceride levels (eg, fasting triglyceride levels) (eg, reducing blood/serum triglyceride levels by at least 20%, 30%, 40%, 50%, 60%, 70%) %, 80%, or 90% or more) comprising administering to a subject in need thereof a dose of about 300 mg once every 4 weeks (or once a month) or once every 3 weeks or once every 2 weeks Subjects are administered a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) subcutaneously. In a certain aspect, provided herein is reducing triglyceride levels (eg, fasting triglyceride levels) (eg, reducing blood/serum triglyceride levels by at least 20%, 30%, 40%, 50%, 60%, 70%) %, 80%, or 90% or more), the method comprising administering to a subject in need thereof a dose in the range of about 200 mg to 250 mg once every 3 weeks or once every 2 weeks or once a week Human FGF21 protein variants (eg, Fc fusion proteins such as V103 (SEQ ID NO: 11)) are administered subcutaneously. In one particular aspect, the methods provided herein are capable of reducing triglyceride levels (eg, fasting triglyceride levels) by at least about 40% or at least about 50% for a period of at least 1-4 weeks. In a certain aspect, provided herein is a method of reducing triglyceride levels (eg, fasting triglyceride levels) by at least about 40% to 60% (eg, at least about 50%) for a period of at least 1-4 weeks, the method comprising A human FGF21 protein variant (eg, an Fc fusion protein such as V103 is administered subcutaneously to a subject in need thereof at a dose of approximately 300 mg once every 4 weeks (or once a month) or once every 3 weeks or once every 2 weeks (SEQ ID NO: 11)). In a certain aspect, provided herein is a method of reducing triglyceride levels (eg, fasting triglyceride levels) by at least about 40% to 60% (eg, at least about 50%) for at least 1-4 weeks, the method comprising with Doses in the range of 200 mg to 300 mg (eg, at least 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, or 280 mg) once every 4 weeks (or monthly) or once every 3 weeks or once every 2 weeks. A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered subcutaneously to a subject in need thereof. In particular aspects, the methods provided herein are for treating human subjects with hypercholesterolemia (eg, primary hypercholesterolemia) or dyslipidemia (eg, mixed dyslipidemia). In certain aspects, the methods provided herein are for treating human subjects suffering from hypertriglyceridemia, particularly severe hypertriglyceridemia. In certain aspects, the methods provided herein are for treating a human subject with type 2 diabetes or an obese human subject. In certain aspects, the methods provided herein are for the treatment of a human subject between 18 and 55 years of age who, when screened prior to administration of a human FGF21 protein variant, has (i ⁾ a 30-45 kg/m BMI within the range (inclusive) (with race-adjusted greater than or equal to 27.5 kg/ ^m2 BMI for Asian subjects), and (ii) within the range of 150 to 500 mg/dL (1.69-5.65 mmol/L) internal triglyceride levels.

In one particular aspect, the methods provided herein are capable of reducing triglyceride levels (eg, fasting triglyceride levels) by at least about 40% or at least about 50%, eg, for a period of at least 1-4 weeks, the methods comprising administering at about 250 mg A human FGF21 protein variant (e.g., an Fc fusion protein, such as V103 (SEQ ID NO: 11)). In a certain aspect, provided herein are methods of reducing triglyceride levels (eg, fasting triglyceride levels) by at least about 40% to 60% (eg, at least about 50%), eg, for a period of at least 1-4 weeks, said The method comprises subcutaneously administering human FGF21 to a subject in need thereof at a dose in the range of about 250 mg to 300 mg (eg, 300 mg) once every 4 weeks (or once a month) or once every 3 weeks or once every 2 weeks Protein variants (eg, Fc fusion proteins such as V103 (SEQ ID NO: 11)).

In certain aspects, provided herein is a reduction in cardiovascular risk (eg, a reduction of at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% or more) ) method comprising at a dose in the range of 100 mg to 600 mg, for example, once daily, once every 2, 3, 4, 5 or 6 days, once weekly, once every 2 weeks, once every 3 weeks, or once every 4 weeks A therapeutically effective amount of a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered to a subject in need thereof once a week (or once a month). In certain aspects, provided herein is a reduction in cardiovascular risk (eg, a reduction of at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% or more) ) method comprising administering to a subject in need thereof at a dose in the range of about 200 mg to 400 mg or 100 mg to 300 mg, for example once weekly, once every 2 weeks, once every 3 weeks or once every 4 weeks Human FGF21 protein variants (eg, Fc fusion proteins such as V103 (SEQ ID NO: 11)). In one particular aspect, provided herein is a reduction in cardiovascular risk (eg, a reduction of at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% or more) multi) method, the method comprising taking about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, Doses of 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg such as once a day, once every 2, 3, 4, 5 or 6 days, once a week, once every 2 weeks, once every 3 weeks, or once every 4 weeks A human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) is administered (or once a month) to a subject in need thereof. In a certain aspect, provided herein is a reduction in cardiovascular risk (eg, a reduction of at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% or more) multi) method comprising subcutaneously administering to a subject in need thereof a human FGF21 protein variant (e.g., an Fc fusion protein such as V103 (SEQ ID NO: 11)). In a certain aspect, provided herein is a reduction in cardiovascular risk (eg, a reduction of at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% or more) A method comprising subcutaneously administering a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11)) to a subject in need thereof at a dose of about 300 mg once every 3 weeks . In a certain aspect, provided herein is a cardiovascular risk reduction (eg, a reduction of at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50%) A method comprising subcutaneously administering a human FGF21 protein variant (eg, an Fc fusion protein such as V103 (SEQ ID NO: 11) to a subject in need thereof once every 3 weeks at a dose in the range of about 200 mg to 250 mg )). In a certain aspect, provided herein is a reduction in cardiovascular risk (eg, a reduction of at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% or more) multi) method comprising subcutaneously administering to a subject in need thereof a human FGF21 protein variant (e.g., an Fc fusion protein such as V103 (SEQ ID NO) at a dose of about 300 mg once every 2 weeks or once a week :11)). In particular aspects, the methods provided herein are for treating a human subject with hypercholesterolemia (eg, primary hypercholesterolemia) or dyslipidemia (eg, mixed dyslipidemia). In certain aspects, the methods provided herein are for treating human subjects with hypertriglyceridemia, particularly severe hypertriglyceridemia. In certain aspects, the methods provided herein are for treating a human subject with type 2 diabetes or an obese human subject. In certain aspects, the methods provided herein are for treating a human subject between the ages of 18 and 55 who, prior to administration of the human FGF21 protein variant, has (i) in the range of 30-45 kg/m ⁽ BMI, inclusive (with race-adjusted greater than or equal to 27.5 kg/m ² BMI for Asian subjects), and (ii) glycerol in the range of 150 to 500 mg/dL (1.69-5.65 mmol/L) Triester levels.

In certain aspects, the methods provided herein are for the treatment of a human subject between 18 and 55 years of age who, when screened prior to administration of a human FGF21 protein variant, has (i ⁾ a 30-45 kg/m BMI within the range (inclusive) (with race-adjusted greater than or equal to 27.5 kg/ ^m2 BMI for Asian subjects), and (ii) within the range of 150 to 500 mg/dL (1.69-5.65 mmol/L) internal triglyceride levels.

In various aspects, the methods provided herein are for treating a human subject at least 18 years old. In other aspects, the methods provided herein are for treating a human subject at least 55 years old. In other aspects, the methods provided herein are for treating a human subject at least 60 years old. In other aspects, the methods provided herein are for treating a human subject at least 65 years old.

In particular aspects, the methods provided herein are for treating human subjects with a BMI in the range of 30 to 45 kg/m ² . In certain aspects, the methods provided herein are used to treat a human subject having a BMI of greater than or equal to ²⁵ kg/m2. In certain aspects, the methods provided herein are for treating a human subject having a BMI of greater ^than or equal to 30 kg/m2. In certain aspects, the methods provided herein are used to treat a human subject having a BMI of greater ^than or equal to 35 kg/m2. In some aspects, the methods provided herein are used to treat human subjects with a BMI (adjusted for ethnicity) > 27.5 for Asian individuals.

In particular aspects, the methods provided herein are for treating human subjects with triglyceride levels in the range of 150-500 mg/dL (1.69-5.65 mmol/L) prior to administration of a human FGF21 protein variant. In particular aspects, the methods provided herein are for treating human subjects with triglyceride levels of at least 150 mg/dL (at least 1.69 mmol/L) prior to administration of a human FGF21 protein variant. In certain aspects, the methods provided herein are for treating human subjects with triglyceride levels of at least 200 mg/dL at screening prior to administration of a human FGF21 protein variant. In particular aspects, the methods provided herein are for treating human subjects with triglyceride levels of at least 500 mg/dL (at least 5.65 mmol/L) prior to administration of a human FGF21 protein variant.

In certain aspects, a human subject with one or more of the following is not treated according to the methods provided herein:

History of hepatobiliary disease, cholelithiasis, or bile sludge (on medical history or on ultrasound screening), hepatic encephalopathy, esophageal varices, or portal shunt;

Liver disease or liver injury as indicated by liver function tests (ALT, AST, GGT, alkaline phosphatase, or serum bilirubin) above the upper limit of normal at screening or baseline;

chronic infection with human immunodeficiency virus (HIV), hepatitis B (HBV) or hepatitis C (HCV);

• A positive HBV surface antigen (HBsAg) test, or under standard local practice, a positive HBV core antigen test.

positive (detectable) HCV RNA;

a history of pancreatic injury or pancreatitis or other pancreatic disease;

Amylase or lipase exceeding ULN at screening or baseline;

History of hypersensitivity reactions to drugs directed against similar biologics, FGF21 protein analogs, or Fc fusion proteins;

a history of bone disorders (including but not limited to osteoporosis, osteopenia, osteomalacia, severe vitamin D deficiency); and

· Plasma 25-hydroxyvitamin D levels below the lower limit of the normal range at screening.

In one aspect, provided herein is a combination therapy for the treatment, prevention or management of a metabolic disorder or a cardiovascular disorder, the combination therapy comprising administering a therapeutically effective amount of a FGF21 protein variant described herein (eg, an Fc fusion protein such as V103) and one or more additional therapeutically active agents (eg, therapeutic agents for metabolic or cardiovascular disorders). Non-limiting examples of other therapeutically active agents for use in combination with the FGF21 protein variants provided herein include obesity treatment agents (eg, phentermine/topiramate, orlistat, lorcaserin, liraglutide, amphetamines) ketone/naltrexone), hypertension therapeutics (eg, diuretics, beta-blockers, alpha-blockers, ACE inhibitors, angiotensin II receptor blockers (ARBs), direct renin inhibition agents, calcium channel blockers, central agonists, peripheral adrenergic blockers, vasodilators, and combinations), diabetes therapeutics (e.g. insulin, alpha-glucosidase inhibitors, biguanides, dopamine agonists, DPP-4 inhibitors, glucagon-like peptides, meglitinides, sodium glucose transporter (SGLT) inhibitors, sulfonylureas, thiazolidinediones, pramlintide), NAFLD/NASH and cardiac Vascular therapeutics (eg, statins, fibrates, aspirin, anticoagulants).

In one aspect, provided herein is a combination therapy for the treatment, prevention or management of a metabolic disorder or a cardiovascular disorder, the combination therapy comprising administering a therapeutically effective amount of a FGF21 protein variant described herein (eg, an Fc fusion protein such as V103) and one or more therapeutically active agents selected from the group consisting of: amiloride (Midamor), bumetanide (Bumex), chlorthalidone (Haigu) Hygroton), chlorothiazide (Diuril), furosemide (Lasix), hydrochlorothiazide or HCTZ (Esidrix, Hydrodiuril, Microzide) ), indapamide (Lozol), metolazone (Mykrox, Zaroxolyn), spironolactone (Aldactone) , Triamterene (Dyrenium), Acebutolol (Sectral), Atenolol (Tenormin), Betaxolol (Kerlone), Bisoprolol (Zebeta), Carteolol (Cartrol), Metoprolol (Lopressor ), Toprol XL), Nadolol (Corgard), Nebivolol (Bystolic), Penbuterol ( Penbutolol (Levatol), Pindolol (Visken), Propranolol (Inderal), Sotalol ( Sotalol (Betapace), Timolol (Timolol (Blocadren)), Doxazosin (Cardura), Prazosin (Pulse) Ningpin (Minipress)), Terazosin (Terazosin) (Hytrin), Benazepril (Lotensin), Captopril (Capt opril) (Capoten), Enalapril (Vasotec), Fosinopril (Monopril), Lisinopril (Cardiac) Prinivil, Zestril), Moexipril (Univasc), Perindopril (Aceon), Quinapril Quinapril (Accupril), Ramipril (Altace), Trandolapril (Mavik), Norvasc (amlodipine), Boi Plendil (felodipine), DynaCirc (isradipine), Cardene (nicardipine), Procardia XL, Adalat (nifedipine), Cardizem, Dilacor, Tiazac, Diltia XL (diltiazem), Sular ( Nisoldipine), Isoptin, Calan, Verelan, Covera-HS (verapamil), Capoten (A Captopril), Vasotec (captopril), Prinivil, Zestril (lisinopril), Lottin (Lotensin) (benazepril), Monopril (fosinopril), Altace (ramipril), Accupril (quinapril ( quinapril), Aceon (perindopril), Mavik (trandolapril), Univasc (moexipril), Candi Atacand (candesartan), irbesartan (Avapro) (irbesartan), olmesartan medoxomil (Benicar) (olmesartan) n)), Cozaar (losartan), Diovan (valsartan), Micardis (telmisartan), Eprosartan ( Teveten) (eprosartan), chlorthalidone (Hygroton), chlorothiazide (Diuril), hydrochlorothiazide or HCTZ (Esidrix, Hydrodiuril, Microzide, indapamide (Lozol), metolazone (Mykrox, Zaroxolyn), amiloride (Midamor), bumetanide (Bumex), furosemide (Lasix), spironolactone (Aldactone), ammonia Triamterene (Dyrenium), Acebutolol (Sectral), Atenolol (Tenormin), Beta Betaxolol (Kerlone), Bisoprolol (Zebeta, Ziac), Carteolol (Cartrol), Carvedilol (Coreg )), Labetalol (Normodyne, Trandate), Metoprolol (Lopressor, Toprol) )-XL), Nadolol (Corgard), Nebivolol (Bystolic), Penbutolol (Penbutolol) (Levatol), Pindolol (Visken), Propanolol (Inderal), Sotalol (Betapace), Timolol (Timolol) (Timolol (Blocadren)), Fibric Acid Derivatives, Hydrochloric Acid, and Omega-3 Fatty Acids, Fenofibrate, Gemfibrozil, Atorvastatin , Fluvastatin, Lovastatin, Pitavastatin, Pravastatin, Rosuvastatin, Simvastatin, Pramlintide, Acarbose (Precose), Miglitol (miglitol) (mignidol (Glyset)), metformin, bromocriptine, alogliptin, linagliptin, saxagliptin, sitagliptin, albiglutide (albiglutide) Tanzeum), dulaglutide (Trulicity), exenatide (Byetta), exenatide extended-release (Xenatide) Senatide (Bydureon)), liraglutide (Victoza), nateglinide (Starlix), repaglinide (Repaglinide) Naphthalene (Prandin), repaglinide-metformin (Prandimet), dapagliflozin (Farxiga), repaglinide-metformin (metformin) ) (Xigduo XR), canagliflozin (Invokana), canagliflozin-metformin (Invokamet), empagliflozin (Jardiance), empagliflozin-linagliptin (Glyxambi), empagliflozin-metformin (Synjardy), sotagliflozin, tofogliflozin , Remogliflozin, luseogliflozin, ipragliflozin, atigliflozin, bexagliflozin, henagliflozin, licogliflozin , glimepiride (Amaryl), glimepiride-pioglitazone (Duetact), glimeperide-rosiglitazone (Avandaryl) ), gliclazide ), glipizide-metformin (Metaglip), glyburide (DiaBeta, Glynase, Micronase), Glyburide-metformin (Glucovance), chlorpropamide (Diabinese), tolazamide (Tolinase), methyl sugar tolbutamide (Orinase, Tol-Tab), rosiglitazone (Avandia), rosiglitazone-glimepiride (Avandaryl) , rosiglitazone-metformin (Amaryl M), pioglitazone (Actos), pioglitazone-alogliptin (Oseni), pioglitazone )-glimepiride (Duetact), and pioglitazone-metformin (Actoplus Met, Actoplus Met XR).

In some embodiments, the sodium glucose transporter (SGLT) inhibitor is selected from the group consisting of dapagliflozin, empagliflozin, canagliflozin, ertugliflozin, Sotagliflozin, Tofogliflozin, Remogliflozin, Luseogliflozin, Ipragliflozin, Atigliflozin, Beglie bexagliflozin, henagliflozin, licogliflozin, and pharmaceutically acceptable salts of any of these. In some embodiments, the sodium glucose transporter (SGLT) inhibitor is dapagliflozin. In some embodiments, the sodium glucose transporter (SGLT) inhibitor is empagliflozin. In some embodiments, the sodium glucose transporter (SGLT) inhibitor is canagliflozin. In some embodiments, the sodium glucose transporter (SGLT) inhibitor is empagliflozin. In some embodiments, the sodium glucose transporter (SGLT) inhibitor is licogliflozin.

In some embodiments, the sodium glucose transporter (SGLT) inhibitor is dapagliflozin or a pharmaceutically acceptable salt thereof. In some embodiments, the sodium glucose transporter (SGLT) inhibitor is empagliflozin or a pharmaceutically acceptable salt thereof. In some embodiments, the sodium glucose transporter (SGLT) inhibitor is canagliflozin or a pharmaceutically acceptable salt thereof. In some embodiments, the sodium glucose transporter (SGLT) inhibitor is ipagliflozin or a pharmaceutically acceptable salt thereof. In some embodiments, the sodium glucose transporter (SGLT) inhibitor is licogliflozin or a pharmaceutically acceptable salt thereof.

In particular aspects, the methods provided herein, comprising administering a FGF21 protein variant (eg, an Fc fusion protein, such as V103), are used as a method of diet (eg, a healthy diet, a calorie-restricted diet), exercise, and/or other lifestyle modifications. Auxiliary.

FGF21 variants

Modifications to FGF21 can improve half-life and/or potency compared to wild-type FGF21 to provide for the treatment or management of metabolic or cardiovascular disorders (e.g., alleviating one or more of the disorders) with clinically preferred dosing regimens. symptoms) or FGF21 therapeutics for cardiovascular risk reduction.

In particular aspects, provided herein are FGF21 protein variants (eg, FGF21 protein variants described in Table 1) for use in the treatment or management of metabolic or cardiovascular disorders (eg, alleviation of one or more symptoms of the disorder) or In methods for reducing cardiovascular risk.

Acceptable amino acid substitutions and modifications that constitute the differences between the FGF21 polypeptides and protein variants and mutants of the methods provided herein and wild-type FGF21 include, but are not limited to, one or more amino acid substitutions (including analogs with non-naturally occurring amino acids) substitution) and truncation. Thus, FGF21 protein variants include, but are not limited to, site-directed FGF21 mutants, truncated FGF21 polypeptides, proteolysis-resistant FGF21 mutants, aggregation-reduced FGF21 mutants, FGF21 combination mutants, FGF21 conjugates (eg, fatty acid- FGF21 conjugates, PEG-FGF21 conjugates) and FGF21 fusion proteins (eg, Fc domain fusion proteins, human serum albumin fusion proteins).

In one aspect, the FGF21 protein variant comprises substitutions of one or more or all of the following residues (according to the numbering scheme of SEQ ID NO: 1) relative to wild-type FGF21: Q55, R105, G148, K150, P158, S195 , P199, and G202.

In one aspect, the FGF21 protein variant comprises one or more or all of the following substitutions (according to the numbering scheme of SEQ ID NO: 1) relative to wild-type FGF21: Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A.

Exemplary fusion protein sequences provided herein are listed in Table 1. Descriptions of such fusions include FGF21 variants and linkers (where applicable). Changes or substitutions employed by FGF21 variants are numbered and described relative to wild-type FGF21.

For example, "Variant 101 (V101)" (SEQ ID NO: 10) is Fc-FGF21 with two amino acid linkers and the following substitutions (according to the numbering scheme of SEQ ID NO: 1) relative to wild-type FGF21 Fusions: Q55C, A109T, G148C, K150R, P158S, P174L, S195A, P199G, and G202A.

By way of another example, "Variant 103 (V103)" (SEQ ID NO: 11) has two amino acid linkers (GS) and the following substitutions relative to wild-type FGF21 (according to the numbering scheme of SEQ ID NO: 1 ) The Fc-FGF21 fusions: Q55C, R105K, G148C, K150R, P158S, S195A, P199G, G202A.

Table 1: Exemplary FGF21 Variants

*- Note, unless otherwise stated, the FGF21 wild-type sequence in this table refers to NCBI reference number NP_061986.1 (SEQ ID NO: 1).

All mutations in the FGF21 portion and their corresponding amino acid numbering references (SEQ ID NO: 1 ) other than those in this table may also include the full-length sequences of the Fc and linker regions.

In particular aspects, provided herein are methods of treating, preventing or managing metabolic or cardiovascular disorders (eg, obesity, NALFD, NASH or diabetes) comprising administering a human FGF21 protein variant comprising A fusion protein of a human Fc region (eg, a modified human Fc region) fused to a mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of: according to SEQ ID NO :1 numbered Q55C, R105K, G148C, K150R, P158S, S195A, P199G and G202A.

In particular aspects, provided herein are methods of treating, preventing or managing metabolic or cardiovascular disorders (eg, obesity, NALFD, NASH or diabetes) comprising administering a human FGF21 protein variant comprising A fusion protein of a human Fc region (eg, a modified human Fc region) fused to the mature human FGF21 protein or fragment thereof, eg, V103 comprising the amino acid sequence of SEQ ID NO:11.

Functional assays for determining or measuring the biological activity of FGF21 or a variant or fragment thereof have been described, eg, see PCT Publication No. WO 2013/049247, which is incorporated herein by reference in its entirety. In certain aspects, the functional assay is an in vitro assay. In other aspects, the functional assay is an in vivo assay. In certain aspects, the functional assay is an ex vivo assay.

In certain aspects, FGF21 downstream biomarkers can be assessed to determine the biological activity of FGF21 or a variant or fragment thereof. As used herein, a "FGF21 downstream biomarker" is a gene or gene product, or a measurable marker of a gene or gene product. In some aspects, the gene or activity that is a downstream marker of FGF21 exhibits altered expression levels, or is expressed in vascular tissue. In some aspects, the activity of the downstream marker is altered in the presence of the FGF21 modulator. In some aspects, the downstream marker exhibits altered expression levels when FGF21 is disrupted by an FGF21 modulator. FGF21 downstream markers include, but are not limited to, glucose or 2-deoxy-glucose uptake, pERK and other phosphorylated or acetylated proteins or NAD levels.

Pharmaceutical compositions and formulations

In particular aspects, provided herein are pharmaceutical compositions comprising FGF21 protein variants (eg, FGF21 protein variants described in Table 1, eg, V103 (SEQ ID NO: 11)) for use in the treatment, prevention and/or management of metabolic disorders or cardiovascular disorders (eg, hypertriglyceridemia and heart disease risk, insulin resistance (eg, in patients with genetic mutations in the insulin receptor) and lipodystrophy, diabetes, obesity, and nonalcoholic fatty liver disease (NAFLD) / nonalcoholic steatohepatitis (NASH)).

The pharmaceutical compositions described herein may comprise a therapeutically effective amount of the FGF21 protein variant and a pharmaceutically or physiologically acceptable carrier. The carrier is generally selected to be suitable for the intended mode of administration, and can include functions for changing, maintaining or maintaining, for example, the pH, osmotic pressure, viscosity, clarity, color, isotonicity, odor, sterility, stability, Dissolution or release rate, adsorption and/or permeation of agents. Typically, these carriers include aqueous or alcoholic/aqueous solutions, emulsions or suspensions, including saline and/or buffered media.

Suitable agents for inclusion in pharmaceutical compositions include, but are not limited to, amino acids (eg, glycine, glutamine, asparagine, arginine, histidine, or lysine), antimicrobial agents, antioxidants (eg, ascorbic acid, sodium sulfite or bisulfite), buffers (such as borate, bicarbonate, Tris-HCl, citrate, phosphate, or other organic acids), bulking agents (such as mannitol or glycine), chelating agents (such as Ethylenediaminetetraacetic acid (EDTA)), complexing agents (such as caffeine, polyvinylpyrrolidone, beta-cyclodextrin or hydroxypropyl-beta-cyclodextrin), bulking agents, monosaccharides, disaccharides and other carbohydrates Compounds (such as glucose, mannose or dextrin), proteins (such as free serum albumin, gelatin, or immunoglobulins), colorants, flavors and diluents, emulsifiers, hydrophilic polymers (such as polyvinylpyrrolidone) ), low molecular weight polypeptides, salt-forming counterions (e.g. sodium), preservatives (e.g. benzalkonium chloride, benzoic acid, salicylic acid, thimerosal, phenethyl alcohol, methylparaben, propylparaben, chlorhexidine, sorbic acid or hydrogen peroxide), solvents (such as glycerol, propylene glycol or polyethylene glycol), sugar alcohols (such as mannitol or sorbitol), suspending agents, surfactants or wetting agents (such as Pluronics; PEG; sorbitan esters; polysorbates (such as polysorbate 20 or polysorbate 80); Triton; tromethamine; lecithin; cholesterol or tyloxacin (tyloxapal), stability enhancers (eg, sucrose or sorbitol), tonicity enhancers (eg, alkali metal halides (eg, sodium chloride or potassium chloride), or mannitol sorbitol), delivery vehicles , diluents, excipients and/or pharmaceutical adjuvants

Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, and lactated Ringer's. Suitable physiologically acceptable thickening agents such as carboxymethyl cellulose, polyvinylpyrrolidone, gelatin and alginates may be included. Intravenous vehicles include fluid and nutrient replenishers and electrolyte replenishers, such as those based on Ringer's dextrose. In some cases, it is preferable to include an agent to adjust the tonicity of the composition, eg, a sugar, polyol such as mannitol, sorbitol, or sodium chloride in the pharmaceutical composition. For example, in many cases it is desirable for the composition to be substantially isotonic. Preservatives and other additives may also be present, such as antimicrobials, antioxidants, chelating agents and inert gases. The exact formulation will depend on the route of administration. Additional relevant principles, methods and components of pharmaceutical formulation are well known. (See, eg, Allen, Loyd V. eds., (2012) Remington's Pharmaceutical Sciences, 22nd Ed., which is incorporated by reference for this purpose)

When parenteral administration is contemplated, pharmaceutical compositions are generally in the form of sterile, pyrogen-free, parenterally acceptable compositions. A particularly suitable vehicle for parenteral injection is a suitably preserved sterile isotonic solution. The pharmaceutical composition may be in the form of a lyophilisate, such as a lyophilisate.

In certain aspects, the pharmaceutical compositions provided herein (eg, compositions comprising FGF21 protein variant V103) are for subcutaneous administration. Suitable formulation components and methods for subcutaneous administration of polypeptide therapeutics (eg, antibodies, fusion proteins, etc.) are known in the art. See, eg, Published US Patent Application No. 2011/0044977 and US Patent No. 8,465,739 and US Patent No. 8,476,239, each of which is incorporated by reference for this purpose. Typically, pharmaceutical compositions for subcutaneous administration contain suitable stabilizers (eg amino acids such as methionine and or carbohydrates such as sucrose), buffers and/or tonicity agents.

In particular aspects, provided herein are pharmaceutical compositions comprising FGF21 protein variants (eg, FGF21 protein variants shown in Table 1, such as V103) as lyophilisates. In particular aspects, a pharmaceutical composition provided herein as a lyophilized formulation comprising a FGF21 protein variant (eg, the FGF21 protein variant shown in Table 1, such as V103) is administered to a subject (eg, administered subcutaneously) ) before reconstitution into solution.

In particular aspects, a pharmaceutical composition provided herein comprising a FGF21 protein variant (eg, the FGF21 protein variant shown in Table 1, such as V103) is in a solution (eg, a FGF21 protein variant such as V103) comprising 100 mg/mL ( For example, after lyophilisate reconstitution). In some aspects, a pharmaceutical composition provided herein comprising a FGF21 protein variant (eg, the FGF21 protein variant shown in Table 1, such as V103) is in a composition comprising at least about 50 mg/mL, 60 mg/mL, 70 mg/mL, 80 mg /mL, 90 mg/mL, 100 mg/mL, 110 mg/mL, 120 mg/mL, 130 mg/mL, 140 mg/mL, or 150 mg/mL of a solution of an FGF21 protein variant (eg, V103) (e.g., at lyophilisate weight) post-construction) form. In some aspects, a pharmaceutical composition provided herein comprising a FGF21 protein variant (eg, the FGF21 protein variant shown in Table 1, such as V103) at a concentration of about 50 mg/mL to 150 mg/mL or 100 mg/mL to 150 mg/mL mL of FGF21 protein variant (eg, V103) in the form of a solution (eg, after lyophilisate reconstitution). In one aspect, a pharmaceutical composition provided herein comprising a FGF21 protein variant (eg, the FGF21 protein variant shown in Table 1, such as V103) is at a concentration comprising about 75 mg/mL to 150 mg/mL of V103 (eg, 100 mg/mL) V103 (SEQ ID NO: 11)) in the form of a solution (eg, after reconstitution of the lyophilisate).

In some aspects, a pharmaceutical composition provided herein comprising a FGF21 protein variant (eg, the FGF21 protein variant shown in Table 1, such as V103) is in a buffer comprising tromethamine (eg, 10 mM, 20 mM, 30 mM, 40 mM) , or 50 mM tromethamine buffer concentration) in the form of a solution (eg, after reconstitution of the lyophilisate). In some aspects, a pharmaceutical composition provided herein comprising a FGF21 protein variant (eg, the FGF21 protein variant shown in Table 1, such as V103) is in a buffer comprising tromethamine (eg, 10 mM to 50 mM tromethamine) alcohol buffer, such as a concentration of 30 mM tromethamine buffer) in the form of a solution (eg, after reconstitution of the lyophilisate).

In some aspects, a pharmaceutical composition provided herein comprising a FGF21 protein variant (eg, the FGF21 protein variant shown in Table 1, such as V103) is in a formulation comprising sucrose (eg, at least about 200 mM, 210 mM, 220 mM, 230 mM, 240 mM, 250 mM, 260 mM, 270 mM, 280 mM, 290 mM, or 300 mM sucrose) in the form of a solution (eg, after lyophilisate reconstitution). In some aspects, a pharmaceutical composition provided herein comprising a FGF21 protein variant (eg, the FGF21 protein variant shown in Table 1, eg, V103) is in a solution comprising sucrose (eg, at least about 250 mM to 300 mM sucrose, eg, 270 mM) (eg, after reconstitution of lyophilisates).

In some aspects, a pharmaceutical composition provided herein comprising a FGF21 protein variant (eg, the FGF21 protein variant shown in Table 1, such as V103) is in a solution comprising a polysorbate (eg, polysorbate 20) ( For example, after lyophilisate reconstitution). In particular aspects, a pharmaceutical composition provided herein comprising a FGF21 protein variant (eg, the FGF21 protein variant shown in Table 1, such as V103) is at a concentration of at least about 0.01%, 0.02%, 0.03%, 0.04%, 0.05% %, 0.06%, 0.07%, 0.08%, 0.09%, or 0.10% polysorbate 20 in solution (eg, after lyophilisate reconstitution). In particular aspects, a pharmaceutical composition provided herein comprising a FGF21 protein variant (eg, the FGF21 protein variant shown in Table 1, such as V103) at a concentration of 0.02% to 0.10% polysorbate 20 (eg, 0.06% polysorbate 20) sorbitan ester 20) in the form of a solution (eg, after reconstitution of the lyophilisate).

In some aspects, a pharmaceutical composition provided herein comprising a FGF21 protein variant (eg, the FGF21 protein variant shown in Table 1, such as V103) is at a pH in the range of 6.5 to 9 (eg, a pH of 8.0) In the form of a solution (eg, after reconstitution of a lyophilisate). In some aspects, a pharmaceutical composition provided herein comprising a FGF21 protein variant (eg, the FGF21 protein variant shown in Table 1, such as V103) is at least about 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0 or in the form of a solution at a pH of 9.5 (eg, after reconstitution of the lyophilisate).

In some aspects, a pharmaceutical composition provided herein comprising a FGF21 protein variant (eg, the FGF21 protein variant shown in Table 1, such as V103) is in a tromethamine buffer, sucrose, and polysorbate 20 In the form of a solution (eg, after reconstitution of a lyophilisate).

In particular aspects, a pharmaceutical composition provided herein comprising a FGF21 protein variant (eg, the FGF21 protein variant shown in Table 1, such as V103) is in a formulation comprising about 80 mg/mL to 100 mg/mL of the FGF21 protein variant (eg, V103 (SEQ ID NO: 11)), approximately 30 mM tromethamine buffer, 270 mM sucrose, and a solution of 0.06% polysorbate 20 (pH 7.5 to 8.5, eg, pH 8.0) (eg, reconstituted in lyophilisate later) form.

In particular aspects, a pharmaceutical composition provided herein comprising a FGF21 protein variant (eg, the FGF21 protein variant shown in Table 1, such as V103) is in a formulation comprising about 100 mg/mL of the FGF21 protein variant (eg, V103 (SEQ ID 1). NO: 11)), approximately 30 mM tromethamine buffer, 270 mM sucrose and 0.06% polysorbate 20 in the form of a solution (eg, after lyophilisate reconstitution) (pH 8.0).

In certain aspects, provided herein are methods of making pharmaceutical compositions comprising a FGF21 protein variant (eg, the FGF21 protein variant shown in Table 1, such as V103), the methods comprising formulating the FGF21 protein variant suitable for Lyophilisate for reconstitution into solution.

In certain aspects, provided herein are methods of making a pharmaceutical composition comprising a FGF21 protein variant (eg, the FGF21 protein variant shown in Table 1, such as V103), the method comprising reconstituting the lyophilisate into a protein as described herein The described solution, for example, comprises up to about 100 mg/mL (e.g., about 50 mg/mL to about 100 mg/mL. FGF21 protein variant (e.g., V103 (SEQ ID NO: 11)), about 30 mM tromethamine buffer, 270 mM A solution of sucrose and 0.06% polysorbate 20 (pH 8.0).

In certain aspects, provided herein are methods of preparing a pharmaceutical composition comprising a FGF21 protein variant (eg, the FGF21 protein variant shown in Table 1, such as V103), the method comprising preparing a FGF21 comprising about 100 mg/mL A solution of protein variant (eg V103 (SEQ ID NO: 11)), approximately 30 mM tromethamine buffer, 270 mM sucrose and 0.06% polysorbate 20 (pH 8.0).

In one particular aspect, the method of preparing a pharmaceutical composition described herein comprises diluting a solution containing a FGF21 protein variant (eg, the FGF21 protein variant shown in Table 1, such as V103) to 10 mg/mL to 90 mg/mL steps within the range of concentrations. In a specific aspect, the solution is diluted with a saline solution (eg, 0.9% saline solution).

V. Examples

The following examples are only intended to illustrate the invention and not to limit it. Abbreviations are used as conventional in the art.

Example 1

Fc-FGF21 variant fusion proteins (eg, V103 comprising SEQ ID NO: 11) have been described, see eg, PCT Publication No. WO 2013/049247. For example, V103 has been reported to induce 2-deoxyglucose turnover through mouse 3T3L1 adipocytes to induce ERK signaling in a cell-based ERK phosphorylation assay and to reduce deficiency in ob/ob mice (as a mouse model of type 2 diabetes mellitus) total plasma glucose, plasma insulin, and body weight in mice with functional leptin. V103 has been reported to have a longer half-life and is thermodynamically more stable, eg as assessed by melting temperature.

The PK profile of V103 in cynomolgus monkeys has been determined in a 2-dose IV and SC injection toxicity study with a 4-week recovery period and a 13-week IV and SC injection toxicity study with a 13-week recovery period, respectively.

The pharmacokinetics of V103 were similar in both studies, with no apparent gender differences. Moderate accumulation of V103 was observed in the 13-week GLP toxicity study (cumulative index 1.15-2.36 for the four dose groups, as measured by Cmax and AUC _0-7d after the first and last dose).

V103 exposure ( _Cmax and AUC) increased approximately dose proportionally over the dose range of 0.3-100 mg/kg. Intravenous pharmacokinetics are bi-exponential and have a linear terminal elimination phase. V103 was slowly absorbed following subcutaneous administration (100 mg/kg) and maximum exposure was observed 1-4 days after SC administration. The terminal elimination phase is not absorption limiting and the estimated subcutaneous bioavailability is 60%-90%. Table 2 below summarizes the mean toxicokinetic parameters calculated for the penultimate (day 85) and final dose (day 92) of the 13-week GLP compliance toxicology study.

Table 2 Mean TK parameters obtained at the penultimate (day 85) and final dose (day 92) of a 13-week GLP compliance toxicity study in cynomolgus monkeys

Day 85, n=6 animals/main group, 0.3 mg/kg, 3.0 mg/kg, 30 mg/kg iv and 30 mg/kg scV103. Day 92, n=4 animals/recovery group, 30 mg/kg iv and 30 mg/kg sc V103.

_Cmax /dose ((μg/mL)/(mg/kg)); AUC0-7d (μg×h/mL); AUC0-7d/dose ((μg×h/mL)/(mg/kg))

The GLP-validated MSD-based PK assay used to determine V103 concentrations in cynomolgus monkey serum quantified biological activity, but also partially truncated/inactive forms of V103.

Consistent with rat pharmacokinetics, HPLC-MS/MS assessments confirmed that in cynomolgus monkeys, the central portion of the FGF21 subdomain of V103 (T _1/2 ~9-12 days) binds and Required for activity, the C-terminal subdomain (T _1/2 about 5-8 days) is more stable. Table 3 below summarizes the pharmacokinetic properties of the biologically active and truncated/inactive forms of V103 as determined in a 13-week GLP compliance toxicology study.

Table 3 In vivo integrity of V103 following weekly IV and SC administration in a 13-week GLP compliance toxicity study in cynomolgus monkeys

TK parameters were calculated from the average TK profile after the last dose (day 92)

*AUC0-90 days (μg×h/mL)

^aAs measured by MSD-based assay

^bAs measured by LC-MS/MS based assay

Anti-V103 antibodies (ADA) have been observed in many animals. In most cases, the ADA response to treatment was associated with accelerated clearance of V103, however, the overall effect on total and sustained exposure to V103 in these studies was small. The selectivity of the ADA response was assessed during the 13-week GLP toxicity study, and no ADA-positive samples cross-reacted with endogenous wild-type FGF21.

Example 2

The objective of the clinical study was to evaluate the safety and tolerability of multiple doses of V103 administered subcutaneously over 3 months in obese subjects. In addition, the study will identify early efficacy signals for various metabolic diseases associated with elevated triglycerides and/or obesity, and whether V103 appears to be suitable for hypertriglyceridemia, obesity and/or non-alcoholic Clinical safety and efficacy profiles of therapeutics for the development of metabolic disease in patients with fatty liver disease.

goal and destination

One or more primary objectives were to evaluate the safety and tolerability of V103 in obese subjects following repeated administration of V103 by subcutaneous (SC) injection over 12 weeks. Endpoints related to this goal include:

Adverse events (AEs)

·vital signs

Liver function tests for aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin (total bile) and alkaline phosphatase (ALP)

24-hour urinary cortisol

· Electrolyte

Secondary goals include:

• To assess the effect of V103 on triglyceride and lipid profiles at 12 weeks of treatment.

• To assess the potential role of V103 on bone biomarkers of resorption, bone formation and the balance of resorption and deposition at 12 weeks of treatment.

• Assess the effect of V103 on body weight at 12 weeks, as measured by body weight, body mass index (BMI) and waist circumference.

Endpoints related to these goals include:

Total cholesterol, LDL-C, HDL-C and triglycerides at 12 weeks (fasting)

Biomarkers of bone resorption at 12 weeks (serum CTX-1, urine NTX-1)

and biomarker bone formation (serum BSAP, P1NP and osteocalcin) at 12 weeks

Body weight, body mass index (BMI), waist circumference and liver fat percentage at 12 weeks as measured by MRI

Other research goals include:

• Evaluation of the pharmacokinetics (PK) of V103 in obese subjects after repeated dosing over 12 weeks.

• Assess the immunogenicity of V103 following repeated SC administration of V103.

• Assess the potential glycemic metabolic effects of V103.

Non-limiting examples of endpoints related to these other study goals include:

PK parameters will be determined, including: AUC _last , _{Cmax and} Tmax;

Fasting blood glucose, insulin, glucagon and C-peptide, and putative estimates of insulin sensitivity and secretion (HOMA) for up to 12 weeks;

·HbA1c, glycated albumin;

adiponectin; and

- Anti-drug antibodies (ADA) pre- and post-dose over 26 weeks.

Study Design :

This study was designed as a non-validating, multicenter, randomized, investigator and subject blinded, placebo-controlled safety study in obese subjects in which V103 or placebo was administered subcutaneously and treated with Repeat dosing for 3 months.

Approximately 60 subjects were randomized (1:1, active agent:placebo) to receive V103 300 mg by subcutaneous (SC) injection (every 4 weeks for three doses) or matching placebo.

The trial consists of three main periods: (1) Screening, lasting 1 to 4 weeks, (2) Subject and Investigator-blinded randomized treatment period, expected to last 12 weeks, and (3) End-of-study evaluation, the last Approximately 120 days after dose of study drug (approximately 6 half-lives).

Group Expansion: After the interim analysis (eg, after approximately 15 subjects in each group reached the end of treatment (Day 84)), lower doses can be assessed if there is evidence of efficacy and additional doses need to be assessed group to inform future research before population expansion.

Group :

The study population will include approximately 60 adult male and female obese subjects between the ages of 18 and 55, inclusive.

Key Inclusion Criteria:

· Male and female subjects between the ages of 18 and 55 (inclusive);

· Body mass index (BMI) in the range of 30 to 45 kg/ ^m2 inclusive, with ethnic adjustment ≥27.5 for Asian individuals (WHO Expert Consultation, 2004, Lancet [The Lancet] ], 363(9403):157-63); BMI=weight (kg)/[height (m)] ² , inclusive; and

• Triglycerides 150-500 mg/dL (1.69-5.65 mmol/L) at screening, inclusive.

Key Exclusion Criteria:

History of hepatobiliary disease, cholelithiasis, or bile sludge (on medical history or on ultrasound screening), hepatic encephalopathy, esophageal varices, or portal shunt;

Liver disease or liver injury as indicated by liver function tests (ALT, AST, GGT, alkaline phosphatase, or serum bilirubin) above the upper limit of normal at screening or baseline;

• Chronic infection with Human Immunodeficiency Virus (HIV), Hepatitis B (HBV) or Hepatitis C (HCV). Subjects were excluded by a positive HBV surface antigen (HBsAg) test, or a positive HBV core antigen test under standard local practice. Subjects with a positive HCV antibody test should have HCV RNA levels measured. Subjects with positive (detectable) HCV RNA should be excluded;

Fasting triglycerides greater than or equal to 500 mg/dL [5.65 mmol/L], or concomitant use of medication for hypertriglyceridemia (fibrates, omega-3 fatty acids, niacin);

History of pancreatic injury or pancreatitis or other pancreatic disease; amylase or lipase above ULN at screening or baseline;

History of hypersensitivity reactions to drugs directed against similar biologics, FGF21 protein analogs, or Fc fusion proteins;

History of bone disorders (including but not limited to osteoporosis, osteopenia, osteomalacia, severe vitamin D deficiency);

Plasma 25-hydroxyvitamin D levels below the lower limit of the normal range at screening;

Contraindications to MRI;

Weight change (self-reported change of more than 5% or self-reported 5 kg during the past 3 months);

Use of the following weight loss medications: Orlistat (Xenical, Alli, lorcaserin (Belviq), phentermine) -topiramate (Qsymia), naltrexone-bupropion (Contrave), or liraglutide (Victoza or Lalu) Peptide (Saxenda) or other glucagon-like peptide-1 (GLP1) receptor agonists (exenatide (Byetta/Bydureon)), lixisenatide lixisenatide (Luxumia), albiglutide (Tanzeum), or dulaglutide (Trulicity or others) ;as well as

Also excluded from a diet, weight loss or exercise program with specific intent to lose weight within 3 months prior to randomization, or any clinical diagnosis of an eating disorder.

Pharmacokinetic assessment: Cmax, Tmax, _Clast , _Tlast , _AUClast , _AUCtau , _AUCinf , T1/2, Vz/F and CL/F from serum concentration-time data, if available.

Efficacy/PD assessment:

· Fasting triglycerides

·weight

· Critical Security Assessment

Monitoring of adverse events and serious adverse events

· Bone biomarkers

·liver function

· Ultrasound

· Pancreatic function

·Pituitary-adrenal function

Other pituitary-endocrine safety monitoring

· Physical examination and vital signs

Monitoring of routine laboratory markers in blood and urine

· ECG

Columbia-Suicide Severity Rating Scale (C-SSRS)

Mechanistic biomarkers and biomarkers of inflammation

Additional Embodiments

In one aspect, provided herein is a method of treating, preventing or managing a metabolic or cardiovascular disorder, the method comprising administering to a subject in need thereof a human FGF21 protein variant at a dose in the range of 100 mg to 600 mg.

In another aspect, provided herein is a method of treating, preventing or managing hypercholesterolemia, mixed dyslipidemia or hypertriglyceridemia, the method comprising administering to a person in need thereof a dose in the range of 100 mg to 600 mg The subject is administered the human FGF21 protein variant.

In another aspect, provided herein is a method of reducing body weight, the method comprising administering to a subject in need thereof a human FGF21 protein variant at a dose ranging from 100 mg to 600 mg.

In another aspect, provided herein is a method of reducing elevated LDL-C, total C, triglycerides and/or Apo B, the method comprising administering to a subject in need thereof a dose in the range of 100 mg to 600 mg were administered human FGF21 protein variants.

In another aspect, provided herein is a method of increasing HDL-C, the method comprising administering to a subject in need thereof a human FGF21 protein variant at a dose ranging from 100 mg to 600 mg.

In another aspect, provided herein is a method of reducing triglyceride levels, the method comprising administering to a subject in need thereof a human FGF21 protein variant at a dose ranging from 100 mg to 600 mg.

In another aspect, provided herein is a method of reducing cardiovascular risk, the method comprising administering to a subject in need thereof a human FGF21 protein variant at a dose ranging from 100 mg to 600 mg.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing, or managing a metabolic disorder or a cardiovascular disorder, the method comprising administering to a dose in the range of 100 mg to 600 mg in need thereof of subjects administered the human FGF21 protein variant.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing, or managing hypercholesterolemia, mixed dyslipidemia, or hypertriglyceridemia, the method comprising at 100 mg to 600 mg A human FGF21 protein variant is administered to a subject in need thereof at a dose within the range of .

In one aspect, provided herein is a human FGF21 protein variant for use in a method of reducing weight, the method comprising administering to a subject in need thereof the human FGF21 protein variant at a dose in the range of 100 mg to 600 mg body.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of lowering elevated LDL-C, total C, triglycerides and/or Apo B, the method comprising in the range of 100 mg to 600 mg A human FGF21 protein variant is administered to a subject in need thereof at a dose of .

In one aspect, provided herein is a human FGF21 protein variant for use in a method of increasing HDL-C, the method comprising administering to a subject in need thereof human FGF21 at a dose ranging from 100 mg to 600 mg protein variant.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of reducing triglyceride levels, the method comprising administering to a subject in need thereof a human FGF21 protein variant at a dose in the range of 100 mg to 600 mg FGF21 protein variants.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of reducing cardiovascular risk, the method comprising administering to a subject in need thereof human FGF21 at a dose in the range of 100 mg to 600 mg protein variant.

In one aspect, provided herein is the use of a human FGF21 protein variant in the manufacture of a medicament for use in a method of treating, preventing or managing a metabolic disorder or a cardiovascular disorder, the method comprising in the range of 100 mg to 600 mg Dosage The human FGF21 protein variant is administered to a subject in need thereof.

In one aspect, provided herein is the use of a human FGF21 protein variant in the manufacture of a medicament for use in a method of treating, preventing or managing hypercholesterolemia, mixed dyslipidemia or hypertriglyceridemia, said The method comprises administering the human FGF21 protein variant to a subject in need thereof at a dose ranging from 100 mg to 600 mg.

In one aspect, provided herein is the use of a human FGF21 protein variant in the manufacture of a medicament for use in a method of reducing body weight, the method comprising administering to a subject in need thereof a dose in the range of 100 mg to 600 mg were administered human FGF21 protein variants.

In one aspect, provided herein is the use of a human FGF21 protein variant in the manufacture of a medicament for use in a method of lowering elevated LDL-C, total C, triglycerides and/or Apo B, the method comprising: The human FGF21 protein variant is administered to a subject in need thereof at doses ranging from 100 mg to 600 mg.

In one aspect, provided herein is the use of a human FGF21 protein variant in the manufacture of a medicament for use in a method of increasing HDL-C, the method comprising administering to a medicament in need thereof in a dose ranging from 100 mg to 600 mg The subject is administered the human FGF21 protein variant.

In one aspect, provided herein is the use of a human FGF21 protein variant in the manufacture of a medicament for use in a method for reducing triglyceride levels, the method comprising administering to a dose in the range of 100 mg to 600 mg in need thereof of subjects administered the human FGF21 protein variant.

In one aspect, provided herein is the use of a human FGF21 protein variant in the manufacture of a medicament for use in a method of reducing cardiovascular risk, the method comprising administering to a person in need thereof a dose in the range of 100 mg to 600 mg The subject is administered the human FGF21 protein variant.

In one aspect, provided herein is the use of a human FGF21 protein variant in the manufacture of a medicament for use in a method of reducing cardiovascular risk, the method comprising administering to a person in need thereof a dose in the range of 100 mg to 600 mg The subject is administered the human FGF21 protein variant.

In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the manufacture of a medicament for use in a method of treating, preventing or managing a metabolic or cardiovascular disorder.

In one aspect, provided herein is a method of preparing a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg for use in the treatment, prevention, or management of hypercholesterolemia, mixed dyslipidemia, or hypertriglyceridemia Use in medicines used in.

In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the manufacture of a medicament for use in a method of weight loss.

In one aspect, provided herein is an amount of a human FGF21 protein variant in the range of 100 mg to 600 mg for use in a method for reducing elevated LDL-C, total C, triglycerides and/or Apo B Use in medicine.

In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the manufacture of a medicament for use in a method of increasing HDL-C.

In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the manufacture of a medicament for use in a method of reducing triglyceride levels.

In one embodiment, the method reduces triglyceride levels by at least about 40% or at least about 50%.

In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the manufacture of a medicament for use in a method of reducing cardiovascular risk.

In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the manufacture of a medicament for use in the treatment, prevention or management of metabolic or cardiovascular disorders.

In one aspect, provided herein is an amount of a human FGF21 protein variant in the range of 100 mg to 600 mg for use in the treatment, prevention or management of hypercholesterolemia, mixed dyslipidemia, or hypertriglyceridemia use in medicines.

In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the manufacture of a medicament for use in weight loss.

In one aspect, provided herein is an amount of a human FGF21 protein variant in the range of 100 mg to 600 mg in the manufacture of a medicament for use in lowering elevated LDL-C, total C, triglycerides and/or Apo B the use of.

In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the manufacture of a medicament for use in increasing HDL-C.

In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the manufacture of a medicament for use in lowering triglycerides.

In one embodiment, the method reduces triglyceride levels by at least about 40% or at least about 50%.

In one aspect, provided herein is the use of a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg in the manufacture of a medicament for use in reducing cardiovascular risk.

In one aspect, provided herein is the use of a human FGF21 protein variant in the manufacture of a medicament for the treatment of a metabolic disorder or a cardiovascular disorder, wherein a unit dose of the human FGF21 protein variant is in the range of 100 mg to 600 mg.

In one aspect, provided herein is the use of a human FGF21 protein variant in the manufacture of a medicament for the treatment of hypercholesterolemia, mixed dyslipidemia or hypertriglyceridemia, wherein a unit dose of the human FGF21 protein variant is in In the range of 100mg to 600mg.

In one aspect, provided herein is the use of a human FGF21 protein variant in the manufacture of a medicament for weight loss, wherein a unit dose of the human FGF21 protein variant is in the range of 100 mg to 600 mg.

In one aspect, provided herein is the use of a human FGF21 protein variant in the manufacture of a medicament for reducing levels of LDL-C, total C, triglycerides and/or Apo B, wherein a unit dose of the human FGF21 protein variant is in In the range of 100mg to 600mg.

In one aspect, provided herein is the use of a human FGF21 protein variant in the manufacture of a medicament for use in a method of increasing HDL-C, wherein a unit dose of the human FGF21 protein variant is in the range of 100 mg to 600 mg.

In one aspect, provided herein is the use of a human FGF21 protein variant in the manufacture of a medicament for reducing triglyceride levels, wherein a unit dose of the human FGF21 protein variant is in the range of 100 mg to 600 mg.

In one aspect, provided herein is the use of a human FGF21 protein variant in the manufacture of a medicament for use in a method of reducing cardiovascular risk, wherein a unit dose of the human FGF21 protein variant is in the range of 100 mg to 600 mg.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing a metabolic disorder or a cardiovascular disorder in a human subject, wherein the human FGF21 protein variant is administered in an amount of 100 mg to 600 mg Doses within the range are provided for administration. In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing a metabolic disorder or a cardiovascular disorder in a human subject, wherein the human FGF21 protein variant is administered in an amount of 100 mg to 600 mg Amounts within the range are provided for administration.

In some embodiments of these aspects, the metabolic disorder or cardiovascular disorder is selected from the group consisting of hypercholesterolemia, dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH) ), type 2 diabetes and obesity.

In some embodiments of these aspects, treating, preventing or managing a metabolic or cardiovascular disorder comprises or is characterized by reducing one or more of the following: a subject's body weight, liver fat content, elevated LDL-C, Total C, triglycerides and Apo B levels. In certain embodiments, treating, preventing or managing a metabolic disorder or cardiovascular disorder comprises or is characterized by increasing HDL-C levels in the subject.

In some embodiments of these aspects, treating, preventing, or managing a metabolic or cardiovascular disorder comprises or is characterized by reducing the subject's triglyceride levels by at least about 40% or at least about 50%. In some embodiments, treating, preventing or managing a metabolic or cardiovascular disorder comprises or is characterized by reducing triglyceride levels in a subject by at least about 40%. In some embodiments, treating, preventing or managing a metabolic or cardiovascular disorder comprises or is characterized by reducing the subject's triglyceride levels by at least about 50%.

In some embodiments of these aspects, treating, preventing, or managing a metabolic or cardiovascular disorder comprises or is characterized by reducing a subject's cardiovascular risk.

In some embodiments of these aspects, the metabolic or cardiovascular disorder is dyslipidemia (optionally mixed dyslipidemia), hypertriglyceridemia (optionally severe hypertriglyceridemia), or hypercholesterolemia Hypercholesterolemia (optionally primary hypercholesterolemia). In some embodiments, the metabolic or cardiovascular disorder is non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH). In certain embodiments, the subject is 18 to 55 years old. In some embodiments, the subject has a body mass index (BMI) in the range of 30 to 45 kg/m ² (inclusive), with race adjusted > 27.5 for subjects of Asian origin or Asian ancestry. In some embodiments, the subject has a body mass index (BMI) in the range of 30 to 45 kg/m ² inclusive, optionally wherein the BMI is > 27.5 for subjects of Asian origin or ancestry. In some embodiments, the subject has triglyceride levels in the range of 150-500 mg/dL (1.69-5.65 mmol/L) when measured prior to administration of the human FGF21 protein variant.

In some embodiments of these aspects, the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of : Q55C, R105K, G148C, K150R, P158S, S195A, P199G and G202A numbered according to SEQ ID NO:1. In certain embodiments, the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In some embodiments of these aspects, the human FGF21 protein variant is provided for administration in a dose of at least 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, or 400 mg. In some embodiments, the human FGF21 protein variant is provided for administration at a dose of about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, or 190 mg, optionally wherein the human FGF21 protein is mutated The body is provided in doses of approximately 100 mg or 150 mg. In some embodiments, the human FGF21 protein variant is provided at a dose of about 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg For administration, optionally wherein the human FGF21 protein variant is provided in a dose of about 200 mg, 250 mg, or 300 mg. In some embodiments, the human FGF21 protein variant is provided for administration in a dose of at least 100 mg. In some embodiments, the human FGF21 protein variant is provided for administration at a dose of about 100 mg. In some embodiments, the human FGF21 protein variant is provided for administration in a dose of at least 150 mg. In some embodiments, the human FGF21 protein variant is provided for administration at a dose of about 150 mg. In some embodiments, the human FGF21 protein variant is provided for administration in a dose of at least 200 mg. In some embodiments, the human FGF21 protein variant is provided for administration at a dose of about 200 mg. In some embodiments, the human FGF21 protein variant is provided for administration in a dose of at least 250 mg. In some embodiments, the human FGF21 protein variant is provided for administration at a dose of about 250 mg. In some embodiments, the human FGF21 protein variant is provided for administration at a dose of 250 mg. In some embodiments, the human FGF21 protein variant is provided for administration in a dose of at least 300 mg. In some embodiments, the human FGF21 protein variant is provided for administration at a dose of about 300 mg. In some embodiments, the human FGF21 protein variant is provided for administration at a dose of 300 mg.

In some embodiments of these aspects, the human FGF21 protein variant is provided for administration in an amount of at least 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, or 400 mg. In some embodiments, the human FGF21 protein variant is provided for administration in an amount of about 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, or 190 mg, optionally wherein the human FGF21 protein is mutated The body is provided in an amount of approximately 100 mg or 150 mg. In some embodiments, the human FGF21 protein variant is provided in an amount of about 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg For administration, optionally wherein the human FGF21 protein variant is provided in an amount of about 200 mg, 250 mg, or 300 mg. In some embodiments, the human FGF21 protein variant is provided for administration in an amount of at least 100 mg. In some embodiments, the human FGF21 protein variant is provided for administration in an amount of about 100 mg. In some embodiments, the human FGF21 protein variant is provided for administration in an amount of at least 150 mg. In some embodiments, the human FGF21 protein variant is provided for administration in an amount of about 150 mg. In some embodiments, the human FGF21 protein variant is provided for administration in an amount of at least 200 mg. In some embodiments, the human FGF21 protein variant is provided for administration in an amount of about 200 mg. In some embodiments, the human FGF21 protein variant is provided for administration in an amount of at least 250 mg. In some embodiments, the human FGF21 protein variant is provided for administration in an amount of about 250 mg. In some embodiments, the human FGF21 protein variant is provided for administration in an amount of 250 mg. In some embodiments, the human FGF21 protein variant is provided for administration in an amount of at least 300 mg. In some embodiments, the human FGF21 protein variant is provided for administration in an amount of about 300 mg. In some embodiments, the human FGF21 protein variant is provided for administration in an amount of 300 mg.

In some embodiments, the human FGF21 protein variant is provided for subcutaneous administration. In some embodiments, the human FGF21 protein variant is provided for administration once a month or once every 4 weeks, once every 3 weeks, once every 2 weeks, or once a week. In some embodiments, the human FGF21 protein variant is provided for administration once a month or once every 4 weeks, once every 3 weeks, once every 2 weeks, or once a week. In some embodiments, the human FGF21 protein variant is provided for subcutaneous administration.

In one aspect, provided herein is a method of treating, preventing or managing a metabolic disorder or a cardiovascular disorder in a human subject, wherein the method comprises administering to the subject a human FGF21 protein variant at a dose in the range of 100 mg to 600 mg . In one aspect, provided herein is a method of treating, preventing or managing a metabolic disorder or a cardiovascular disorder in a human subject, wherein the method comprises administering to the subject a human FGF21 protein variant in an amount ranging from 100 mg to 600 mg .

In some embodiments, the metabolic disorder or cardiovascular disorder is selected from hypercholesterolemia, dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), 2 type diabetes and obesity.

In some embodiments, treating, preventing or managing a metabolic or cardiovascular disorder comprises or is characterized by reducing one or more of the following: a subject's body weight, liver fat content, elevated LDL-C, total C, Triglyceride and Apo B levels. In certain embodiments, treating, preventing or managing a metabolic disorder or cardiovascular disorder comprises or is characterized by increasing HDL-C levels in the subject.

In some embodiments, treating, preventing or managing a metabolic or cardiovascular disorder comprises or is characterized by reducing the subject's triglyceride levels by at least about 40% or at least about 50%. In some embodiments, treating, preventing or managing a metabolic or cardiovascular disorder comprises or is characterized by reducing triglyceride levels in a subject by at least about 40%. In some embodiments, treating, preventing or managing a metabolic or cardiovascular disorder comprises or is characterized by reducing the subject's triglyceride levels by at least about 50%.

In some embodiments, treating, preventing or managing a metabolic or cardiovascular disorder comprises or is characterized by reducing a subject's cardiovascular risk.

In some embodiments, the metabolic or cardiovascular disorder is dyslipidemia, hypertriglyceridemia, or hypercholesterolemia. In some embodiments, the metabolic disorder or cardiovascular disorder is dyslipidemia. In some embodiments, the metabolic disorder or cardiovascular disorder is hypertriglyceridemia. In some embodiments, the metabolic disorder or cardiovascular disorder is hypercholesterolemia. In some embodiments, the metabolic disorder or cardiovascular disorder is mixed dyslipidemia. In some embodiments, the metabolic disorder or cardiovascular disorder is severe hypertriglyceridemia. In some embodiments, the metabolic disorder or cardiovascular disorder is primary hypercholesterolemia. In some embodiments, the metabolic or cardiovascular disorder is non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH). In some embodiments, the metabolic or cardiovascular disorder is non-alcoholic fatty liver disease (NAFLD). In some embodiments, the metabolic disorder or cardiovascular disorder is nonalcoholic steatohepatitis (NASH). In certain embodiments, the subject is 18 to 55 years old. In some embodiments, the subject has a body mass index (BMI) in the range of 30 to 45 kg/m ² (inclusive), with race adjusted > 27.5 for subjects of Asian origin or Asian ancestry. In some embodiments, the subject has a body mass index (BMI)

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing hypercholesterolemia in a human subject, wherein the human FGF21 protein variant is administered at 100 mg, 110 mg, 120 mg, Within the range of 130mg, 140mg, 150mg, 160mg, 170mg, 180mg, 190mg, 200mg, 210mg, 220mg, 230mg, 240mg, 250mg, 260mg, 270mg, 280mg, 290mg, 300mg, 310mg, 320mg, 330mg, 340mg, or 350mg A dose or amount is provided for administration, optionally wherein the human FGF21 protein variant is provided in a dose of about 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg, and wherein the human FGF21 variant is comprised of a combination with mature human FGF21 protein or A fusion protein of a human Fc region fused to a fragment thereof, the mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, numbered according to SEQ ID NO: 1, P199G, and G202A, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11. In one embodiment thereof, the human FGF21 variant is provided for administration at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg once per week. In one embodiment thereof, the human FGF21 variant is provided for administration once every two weeks at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg. In one embodiment thereof, the human FGF21 variant is provided for administration once every three weeks at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg. In one embodiment thereof, the human FGF21 variant is provided for administration at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg once every four weeks or once a month.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing dyslipidemia in a human subject, wherein the human FGF21 protein variant is administered at 100 mg, 110 mg, 120 mg, 130 mg, Doses or doses in the range of 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg An amount is provided for administration, optionally wherein the human FGF21 protein variant is provided in a dose of about 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg, and wherein the human FGF21 variant is comprised of a mature human FGF21 protein or fragment thereof A fusion protein of a fused human Fc region, the mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G numbered according to SEQ ID NO: 1 , and G202A, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11. In one embodiment thereof, the human FGF21 variant is provided for administration at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg once per week. In one embodiment thereof, the human FGF21 variant is provided for administration once every two weeks at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg. In one embodiment thereof, the human FGF21 variant is provided for administration once every three weeks at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg. In one embodiment thereof, the human FGF21 variant is provided for administration at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg once every four weeks or once a month.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing hypertriglyceridemia in a human subject, wherein the human FGF21 protein variant is administered at 100 mg, 110 mg, 120mg, 130mg, 140mg, 150mg, 160mg, 170mg, 180mg, 190mg, 200mg, 210mg, 220mg, 230mg, 240mg, 250mg, 260mg, 270mg, 280mg, 290mg, 300mg, 310mg, 320mg, 330mg, 340mg, or 350mg range The dose or amount within is provided for administration, optionally wherein the human FGF21 protein variant is provided in a dose of about 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg, and wherein the human FGF21 variant is comprised of mature human FGF21 A fusion protein of a human Fc region fused to a protein or fragment thereof, the mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, numbered according to SEQ ID NO: 1, S195A, P199G, and G202A, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11. In one embodiment thereof, the human FGF21 variant is provided for administration at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg once per week. In one embodiment thereof, the human FGF21 variant is provided for administration once every two weeks at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg. In one embodiment thereof, the human FGF21 variant is provided for administration once every three weeks at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg. In one embodiment thereof, the human FGF21 variant is provided for administration at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg once every four weeks or once a month.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) in a human subject, wherein the human FGF21 protein variant is administered at 100 mg or A dose or amount in the range of 350 mg is provided for administration, optionally wherein the human FGF21 protein variant is provided in a dose of about 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg, and wherein the human FGF21 variant is comprised of A fusion protein of a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, numbered according to SEQ ID NO: 1, K150R, P158S, S195A, P199G, and G202A, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11. In one embodiment thereof, the human FGF21 variant is provided for administration at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg once per week. In one embodiment thereof, the human FGF21 variant is provided for administration once every two weeks at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg. In one embodiment thereof, the human FGF21 variant is provided for administration once every three weeks at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg. In one embodiment thereof, the human FGF21 variant is provided for administration at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg once every four weeks or once a month.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing nonalcoholic steatohepatitis (NASH) in a human subject, wherein the human FGF21 protein variant is administered at 100 mg or A dose or amount in the range of 350 mg is provided for administration, optionally wherein the human FGF21 protein variant is provided in a dose of about 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg, and wherein the human FGF21 variant is comprised of A fusion protein of a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, numbered according to SEQ ID NO: 1, K150R, P158S, S195A, P199G, and G202A, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11. In one embodiment thereof, the human FGF21 variant is provided for administration at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg once per week. In one embodiment thereof, the human FGF21 variant is provided for administration once every two weeks at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg. In one embodiment thereof, the human FGF21 variant is provided for administration once every three weeks at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg. In one embodiment thereof, the human FGF21 variant is provided for administration at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg once every four weeks or once a month.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing type 2 diabetes in a human subject, wherein the human FGF21 protein variant is administered at 100 mg, 110 mg, 120 mg, 130 mg , 140mg, 150mg, 160mg, 170mg, 180mg, 190mg, 200mg, 210mg, 220mg, 230mg, 240mg, 250mg, 260mg, 270mg, 280mg, 290mg, 300mg, 310mg, 320mg, 330mg, 340mg, or 350mg range or an amount is provided for administration, optionally wherein the human FGF21 protein variant is provided in a dose of about 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg, and wherein the human FGF21 variant is a protein comprising mature human FGF21 protein or A fusion protein of a fragment-fused human Fc region, the mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, numbered according to SEQ ID NO: 1, P199G, and G202A, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11. In one embodiment thereof, the human FGF21 variant is provided for administration at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg once per week. In one embodiment thereof, the human FGF21 variant is provided for administration once every two weeks at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg. In one embodiment thereof, the human FGF21 variant is provided for administration once every three weeks at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg. In one embodiment thereof, the human FGF21 variant is provided for administration at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg once every four weeks or once a month.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing obesity in a human subject, wherein the human FGF21 protein variant is administered at 100 mg, 110 mg, 120 mg, 130 mg, Doses or doses in the range of 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, 300 mg, 310 mg, 320 mg, 330 mg, 340 mg, or 350 mg An amount is provided for administration, optionally wherein the human FGF21 protein variant is provided in a dose of about 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg, and wherein the human FGF21 variant is comprised of a mature human FGF21 protein or fragment thereof A fusion protein of a fused human Fc region, the mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G numbered according to SEQ ID NO: 1 , and G202A, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11. In one embodiment thereof, the human FGF21 variant is provided for administration at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg once per week. In one embodiment thereof, the human FGF21 variant is provided for administration once every two weeks at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg. In one embodiment thereof, the human FGF21 variant is provided for administration once every three weeks at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg. In one embodiment thereof, the human FGF21 variant is provided for administration at a dose of about 100 mg, 150 mg, 200 mg, 250 mg or 300 mg once every four weeks or once a month.

In one aspect, provided herein is for use in the treatment, prevention or management of hypercholesterolemia, dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease (NAFLD), nonalcoholic fatty liver disease in a human subject A human FGF21 protein variant for use in a method of hepatitis (NASH), type 2 diabetes or obesity, wherein the human FGF21 protein variant is provided for administration in a dose or amount in the range of 100 mg, and wherein the human FGF21 variant The antibody is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of Q55C, numbered according to SEQ ID NO: 1, R105K, G148C, K150R, P158S, S195A, P199G, and G202A, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11. In one embodiment thereof, the human FGF21 variant is provided for once-weekly administration. In one embodiment thereof, the human FGF21 variant is provided for biweekly administration. In one embodiment thereof, the human FGF21 variant is provided for administration once every three weeks. In one embodiment thereof, the human FGF21 variant is provided for administration every four weeks or monthly.

In one aspect, provided herein is for use in the treatment, prevention or management of hypercholesterolemia, dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease (NAFLD), nonalcoholic fatty liver disease in a human subject A human FGF21 protein variant for use in a method of hepatitis (NASH), type 2 diabetes or obesity, wherein the human FGF21 protein variant is provided for administration in a dose or amount in the range of 150 mg, and wherein the human FGF21 variant The antibody is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of Q55C, numbered according to SEQ ID NO: 1, R105K, G148C, K150R, P158S, S195A, P199G, and G202A, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11. In one embodiment thereof, the human FGF21 variant is provided for once-weekly administration. In one embodiment thereof, the human FGF21 variant is provided for biweekly administration. In one embodiment thereof, the human FGF21 variant is provided for administration once every three weeks. In one embodiment thereof, the human FGF21 variant is provided for administration every four weeks or monthly.

In one aspect, provided herein is for use in the treatment, prevention or management of hypercholesterolemia, dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease (NAFLD), nonalcoholic fatty liver disease in a human subject A human FGF21 protein variant for use in a method of hepatitis (NASH), type 2 diabetes or obesity, wherein the human FGF21 protein variant is provided for administration in a dose or amount in the range of 200 mg, and wherein the human FGF21 variant The antibody is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of Q55C, numbered according to SEQ ID NO: 1, R105K, G148C, K150R, P158S, S195A, P199G, and G202A, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11. In one embodiment thereof, the human FGF21 variant is provided for once-weekly administration. In one embodiment thereof, the human FGF21 variant is provided for biweekly administration. In one embodiment thereof, the human FGF21 variant is provided for administration once every three weeks. In one embodiment thereof, the human FGF21 variant is provided for administration every four weeks or monthly.

In one aspect, provided herein is for use in the treatment, prevention or management of hypercholesterolemia, dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease (NAFLD), nonalcoholic fatty liver disease in a human subject A human FGF21 protein variant for use in a method of hepatitis (NASH), type 2 diabetes or obesity, wherein the human FGF21 protein variant is provided for administration in a dose or amount in the range of 250 mg, and wherein the human FGF21 variant The antibody is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of Q55C, numbered according to SEQ ID NO: 1, R105K, G148C, K150R, P158S, S195A, P199G, and G202A, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11. In one embodiment thereof, the human FGF21 variant is provided for once-weekly administration. In one embodiment thereof, the human FGF21 variant is provided for biweekly administration. In one embodiment thereof, the human FGF21 variant is provided for administration once every three weeks. In one embodiment thereof, the human FGF21 variant is provided for administration every four weeks or monthly.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic steatohepatitis (NASH) in a human subject, wherein the human FGF21 protein variant is administered at 300 mg A dose or amount within the range of the human FGF21 variant is provided for administration, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising a protein selected from the group consisting of One or more mutations of: Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: 11 amino acid sequence. In one embodiment thereof, the human FGF21 variant is provided for once-weekly administration. In one embodiment thereof, the human FGF21 variant is provided for biweekly administration. In one embodiment thereof, the human FGF21 variant is provided for administration once every three weeks. In one embodiment thereof, the human FGF21 variant is provided for administration every four weeks or monthly.

In one aspect, provided herein is for use in the treatment, prevention or management of hypercholesterolemia, dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease (NAFLD), nonalcoholic fatty liver disease in a human subject A human FGF21 protein variant for use in a method of hepatitis (NASH), type 2 diabetes or obesity, wherein the human FGF21 protein variant is provided for administration once a week in a dose or amount in the range of 100 mg or 150 mg, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of: according to SEQ ID NO: 1 Numbered Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is for use in the treatment, prevention or management of hypercholesterolemia, dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease (NAFLD), nonalcoholic fatty liver disease in a human subject a human FGF21 protein variant for use in a method of hepatitis (NASH), type 2 diabetes or obesity, wherein the human FGF21 protein variant is provided for administration once every two weeks in a dose or amount in the range of 150 mg or 200 mg, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of: according to SEQ ID NO: 1 numbered Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is for use in the treatment, prevention or management of hypercholesterolemia, dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease (NAFLD), nonalcoholic fatty liver disease in a human subject a human FGF21 protein variant for use in a method of hepatitis (NASH), type 2 diabetes or obesity, wherein the human FGF21 protein variant is provided for administration once every three weeks in a dose or amount in the range of 200 mg or 250 mg, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of: according to SEQ ID NO: 1 numbered Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is for use in the treatment, prevention or management of hypercholesterolemia, dyslipidemia, hypertriglyceridemia, nonalcoholic fatty liver disease (NAFLD), nonalcoholic fatty liver disease in a human subject A human FGF21 protein variant for use in a method for hepatitis (NASH), type 2 diabetes or obesity, wherein the human FGF21 protein variant is provided every four weeks or monthly in a dose or amount in the range of 250 mg or 300 mg for administration, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of: according to SEQ ID NO:1 numbered Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing hypercholesterolemia in a human subject, wherein the human FGF21 protein variant is provided for use in combination with one or Multiple additional therapeutically active agents are administered in combination, wherein the one or more additional therapeutically active agents are SGLT inhibitors selected from the group consisting of: dapagliflozin, empagliflozin, canagar Ligagliflozin, ipagliflozin, soxagliflozin, topagliflozin, repagliflozin, lupagliflozin, ipagliflozin, apagliflozin, beagliflozin, henggliflozin, licogliflozin and A pharmaceutically acceptable salt of any of these, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one selected from the group consisting of or multiple mutations: Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO:1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11 . In one embodiment thereof, the SGLT inhibitor is licogliflozin.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing dyslipidemia in a human subject, wherein the human FGF21 protein variant is provided for use in combination with one or more Additional therapeutically active agents are administered in combination, wherein the one or more additional therapeutically active agents are SGLT inhibitors selected from the group consisting of dapagliflozin, empagliflozin, canagliflozin , ipagliflozin, soxagliflozin, topagliflozin, regagliflozin, lupagliflozin, ipagliflozin, apagliflozin, bepagliflozin, henggliflozin, licogliflozin and any of these and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one or more selected from the group consisting of Mutations: Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO:1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11. In one embodiment thereof, the SGLT inhibitor is licogliflozin.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing hypertriglyceridemia in a human subject, wherein the human FGF21 protein variant is provided for use in combination with a One or more other therapeutically active agents are used in combination, wherein the one or more other therapeutically active agents are SGLT inhibitors selected from the group consisting of: dapagliflozin, empagliflozin, Canagliflozin, Epagliflozin, Soxagliflozin, Toagliflozin, Regagliflozin, Lupagliflozin, Ipagliflozin, Agliflozin, Beagliflozin, Henggliflozin, licogliflozin and pharmaceutically acceptable salts of any of these, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising a protein selected from the group consisting of One or more mutations of: Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: 11 amino acid sequence. In one embodiment thereof, the SGLT inhibitor is licogliflozin.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) in a human subject, wherein the human FGF21 protein variant is provided with Use in combination with one or more other therapeutically active agents, wherein the one or more other therapeutically active agents are SGLT inhibitors selected from the group consisting of: Dapagliflozin, Engel Ligagliflozin, Canagliflozin, Agliflozin, Soxagliflozin, Toagliflozin, Regagliflozin, Lupagliflozin, Iggliflozin, Agliflozin, Beagliflozin, Hengagliflozin Liekin, licogliflozin and pharmaceutically acceptable salts of any of these, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising a mature human FGF21 protein or fragment thereof One or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO :11 amino acid sequence. In one embodiment thereof, the SGLT inhibitor is licogliflozin.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic steatohepatitis (NASH) in a human subject, wherein the human FGF21 protein variant is provided for use with Use in combination with one or more other therapeutically active agents, wherein the one or more other therapeutically active agents are SGLT inhibitors selected from the group consisting of: Dapagliflozin, Engel Ligagliflozin, Canagliflozin, Agliflozin, Soxagliflozin, Toagliflozin, Regagliflozin, Lupagliflozin, Iggliflozin, Agliflozin, Beagliflozin, Hengagliflozin Liekin, licogliflozin and pharmaceutically acceptable salts of any of these, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising a mature human FGF21 protein or fragment thereof One or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO :11 amino acid sequence. In one embodiment thereof, the SGLT inhibitor is licogliflozin.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing type 2 diabetes in a human subject, wherein the human FGF21 protein variant is provided for use in combination with one or more A kind of other therapeutically active agent combination administration, wherein this one or more other therapeutically active agent is the SGLT inhibitor selected from the following group, and this group is made up of following: dapagliflozin, empagliflozin, canagliflozin net, ipagliflozin, soxagliflozin, topagliflozin, repagliflozin, lupagliflozin, ipagliflozin, apagliflozin, bepagliflozin, henggliflozin, licogliflozin and any A pharmaceutically acceptable salt of these, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one selected from the group consisting of or Multiple mutations: Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO:1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11. In one embodiment thereof, the SGLT inhibitor is licogliflozin.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing obesity in a human subject, wherein the human FGF21 protein variant is provided for use in combination with one or more Additional therapeutically active agents are administered in combination, wherein the one or more additional therapeutically active agents are SGLT inhibitors selected from the group consisting of dapagliflozin, empagliflozin, canagliflozin , ipagliflozin, soxagliflozin, topagliflozin, regagliflozin, lupagliflozin, ipagliflozin, apagliflozin, bepagliflozin, henggliflozin, licogliflozin and any of these and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one or more selected from the group consisting of Mutations: Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO:1, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11. In one embodiment thereof, the SGLT inhibitor is licogliflozin.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 The protein variant is provided for administration once a week in a dose or amount in the range of 100 mg, and wherein the subject's liver fat content is reduced to within a manageable or normal range as determined by a clinician or clinical guidelines or reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof, the The mature human FGF21 protein or fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human The FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 The protein variant is provided for administration in a dose or amount in the range of 100 mg once every two weeks, and wherein the subject's liver fat content is reduced to within a manageable or normal range or reduced as determined by a clinician or clinical guidelines at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to the mature human FGF21 protein or fragment thereof, The mature human FGF21 protein or fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the The human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 The protein variant is provided for administration in a dose or amount in the range of 100 mg once every three weeks, and wherein the subject's liver fat content is reduced to a manageable or normal range or reduced as determined by a clinician or clinical guidelines at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to the mature human FGF21 protein or fragment thereof, The mature human FGF21 protein or fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the The human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 The protein variant is provided for administration once every four weeks in a dose or amount in the range of 100 mg, and wherein the subject's liver fat content is reduced to within a manageable or normal range as determined by a clinician or clinical guidelines or reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof, the The mature human FGF21 protein or fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human The FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 The protein variant is provided for administration once a week in a dose or amount in the range of 150 mg, and wherein the subject's liver fat content is reduced to within a manageable or normal range as determined by a clinician or clinical guidelines or reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof, the The mature human FGF21 protein or fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human The FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 The protein variant is provided for administration in a dose or amount in the range of 150 mg once every two weeks, and wherein the subject's liver fat content is reduced to a manageable or normal range or reduced as determined by a clinician or clinical guidelines at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to the mature human FGF21 protein or fragment thereof, The mature human FGF21 protein or fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the The human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 The protein variant is provided for administration in a dose or amount in the range of 150 mg once every three weeks, and wherein the subject's liver fat content is reduced to within the manageable or normal range or reduced as determined by the clinician or clinical guidelines at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to the mature human FGF21 protein or fragment thereof, The mature human FGF21 protein or fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the The human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 The protein variant is provided for administration once every four weeks in a dose or amount in the range of 150 mg, and wherein the subject's liver fat content is reduced to within a manageable or normal range as determined by a clinician or clinical guidelines or reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof, the The mature human FGF21 protein or fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human The FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 The protein variant is provided for administration once a week in a dose or amount in the range of 200 mg, and wherein the subject's liver fat content is reduced to within a manageable or normal range as determined by a clinician or clinical guidelines or reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof, the The mature human FGF21 protein or fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human The FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 The protein variant is provided for administration in a dose or amount in the range of 200 mg once every two weeks, and wherein the subject's liver fat content is reduced to a manageable or normal range or reduced as determined by a clinician or clinical guidelines at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to the mature human FGF21 protein or fragment thereof, The mature human FGF21 protein or fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the The human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 The protein variant is provided for administration once every three weeks in a dose or amount in the range of 200 mg, and wherein the subject's liver fat content is reduced to a manageable or normal range or reduced as determined by a clinician or clinical guidelines at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to the mature human FGF21 protein or fragment thereof, The mature human FGF21 protein or fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the The human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 The protein variant is provided for administration once every four weeks in a dose or amount in the range of 200 mg, and wherein the subject's liver fat content is reduced to within a manageable or normal range as determined by a clinician or clinical guidelines or reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof, the The mature human FGF21 protein or fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human The FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 The protein variant is provided for administration once a week in a dose or amount in the range of 250 mg, and wherein the subject's liver fat content is reduced to within a manageable or normal range as determined by a clinician or clinical guidelines or reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof, the The mature human FGF21 protein or fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human The FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 The protein variant is provided for administration in a dose or amount in the range of 250 mg once every two weeks, and wherein the subject's liver fat content is reduced to a manageable or normal range or reduced as determined by a clinician or clinical guidelines at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to the mature human FGF21 protein or fragment thereof, The mature human FGF21 protein or fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the The human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 The protein variant is provided for administration once every three weeks in a dose or amount in the range of 250 mg, and wherein the subject's liver fat content is reduced to a manageable or normal range or reduced as determined by a clinician or clinical guidelines at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to the mature human FGF21 protein or fragment thereof, The mature human FGF21 protein or fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the The human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 The protein variant is provided for administration once every four weeks in a dose or amount in the range of 250 mg, and wherein the subject's liver fat content is reduced to within the manageable or normal range as determined by a clinician or clinical guidelines or reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof, the The mature human FGF21 protein or fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human The FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 The protein variant is provided for administration once a week in a dose or amount in the range of 300 mg, and wherein the subject's liver fat content is reduced to within a manageable or normal range as determined by a clinician or clinical guidelines or reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof, the The mature human FGF21 protein or fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human The FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 The protein variant is provided for administration in a dose or amount in the range of 300 mg once every two weeks and wherein the subject's liver fat content is reduced to a manageable or normal range or reduced as determined by a clinician or clinical guidelines at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to the mature human FGF21 protein or fragment thereof, The mature human FGF21 protein or fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the The human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 The protein variant is provided for administration once every three weeks in a dose or amount in the range of 300 mg, and wherein the subject's liver fat content is reduced to a manageable or normal range or reduced as determined by a clinician or clinical guidelines at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to the mature human FGF21 protein or fragment thereof, The mature human FGF21 protein or fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the The human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH), wherein the human FGF21 The protein variant is provided for administration once every four weeks in a dose or amount in the range of 300 mg, and wherein the subject's liver fat content is reduced to within a manageable or normal range as determined by a clinician or clinical guidelines or reduced by at least about 5%, 10%, 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof, the The mature human FGF21 protein or fragment thereof comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human The FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic fatty liver disease (NAFLD) in a human subject, wherein the human FGF21 protein variant is provided with In combination with licogliflozin or a pharmaceutically acceptable salt thereof, and wherein the subject's liver fat content is reduced to within the manageable or normal range as determined by a clinician or clinical guidelines or by at least about 5%, 10% , 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof, the mature human FGF21 protein or The fragment comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: 1 Amino acid sequence of ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing non-alcoholic steatohepatitis (NASH) in a human subject, wherein the human FGF21 protein variant is provided for use with In combination with licogliflozin or a pharmaceutically acceptable salt thereof, and wherein the subject's liver fat content is reduced to within the manageable or normal range as determined by a clinician or clinical guidelines or by at least about 5%, 10% , 20%, 30%, 40%, or 50% or more, and wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof, the mature human FGF21 protein or The fragment comprises one or more mutations selected from the group consisting of Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A numbered according to SEQ ID NO: 1, optionally wherein the human FGF21 protein variant comprises SEQ ID NO: 1 Amino acid sequence of ID NO: 11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing triglyceridemia, wherein the human FGF21 protein variant is administered in a dose or amount in the range of 100 mg per week provided for administration once, once every two weeks, once every three weeks, once every four weeks, or once a month, and wherein the subject's triglyceride levels are reduced by at least about 40% or at least about 50%, and wherein the person FGF21 variants are fusion proteins comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of: numbered according to SEQ ID NO: 1 Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing triglycerideemia, wherein the human FGF21 protein variant is administered in a dose or amount in the range of 150 mg per week provided for administration once, once every two weeks, once every three weeks, once every four weeks, or once a month, and wherein the subject's triglyceride levels are reduced by at least about 40% or at least about 50%, and wherein the person FGF21 variants are fusion proteins comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of: numbered according to SEQ ID NO: 1 Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing triglycerideemia, wherein the human FGF21 protein variant is administered in a dose or amount in the range of 200 mg per week provided for administration once, once every two weeks, once every three weeks, once every four weeks, or once a month, and wherein the subject's triglyceride levels are reduced by at least about 40% or at least about 50%, and wherein the person FGF21 variants are fusion proteins comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of: numbered according to SEQ ID NO: 1 Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing triglycerideemia, wherein the human FGF21 protein variant is administered in a dose or amount in the range of 250 mg per week provided for administration once, once every two weeks, once every three weeks, once every four weeks, or once a month, and wherein the subject's triglyceride levels are reduced by at least about 40% or at least about 50%, and wherein the person FGF21 variants are fusion proteins comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of: numbered according to SEQ ID NO: 1 Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

In one aspect, provided herein is a human FGF21 protein variant for use in a method of treating, preventing or managing triglycerideemia, wherein the human FGF21 protein variant is administered in a dose or amount in the range of 300 mg per week provided for administration once, once every two weeks, once every three weeks, once every four weeks, or once a month, and wherein the subject's triglyceride levels are reduced by at least about 40% or at least about 50%, and wherein the person FGF21 variants are fusion proteins comprising a human Fc region fused to a mature human FGF21 protein or fragment thereof comprising one or more mutations selected from the group consisting of: numbered according to SEQ ID NO: 1 Q55C, R105K, G148C, K150R, P158S, S195A, P199G, and G202A, optionally wherein the human FGF21 protein variant comprises the amino acid sequence of SEQ ID NO:11.

Each of the foregoing aspects and embodiments, as well as other elements described herein, may be combined in any manner and without limitation.

Claims (20)

1. A human FGF21 protein variant for use in a method of treating, preventing or managing a metabolic or cardiovascular disorder in a human subject, wherein the human FGF21 protein variant is provided for administration at a dose in the range of 100mg to 600 mg.

2. The variant human FGF21 protein for use according to claim 1, wherein the metabolic or cardiovascular disorder is selected from hypercholesterolemia, dyslipidemia, hypertriglyceridemia, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), type 2 diabetes and obesity.

3. The variant human FGF21 protein for use according to claim 1 or 2, wherein treating, preventing or managing the metabolic or cardiovascular disorder comprises or is characterized by decreasing one or more of: subject weight, liver fat content, elevated LDL-C, total C, triglycerides and Apo B levels.

4. The variant human FGF21 protein for use according to any one of claims 1-3, wherein treating, preventing or managing the metabolic or cardiovascular disorder comprises or is characterized by increasing HDL-C levels in a subject.

5. The variant human FGF21 protein for use according to claim 3, wherein treating, preventing or managing the metabolic or cardiovascular disorder comprises or is characterized by reducing triglyceride levels in the subject by at least about 40% or by at least about 50%.

6. The variant human FGF21 protein for use according to claim 1 or 2, wherein treating, preventing or managing the metabolic or cardiovascular disorder comprises or is characterized by reducing the cardiovascular risk of the subject.

7. The FGF21 protein variant for use according to any one of claims 2 to 6, wherein the metabolic or cardiovascular disorder is dyslipidemia, optionally mixed dyslipidemia, hypertriglyceridemia, optionally severe hypertriglyceridemia, or hypercholesterolemia, optionally primary hypercholesterolemia.

8. The FGF21 protein variant for use according to any one of claims 2-6, wherein the metabolic or cardiovascular disorder is non-alcoholic fatty liver disease (NAFLD) or non-alcoholic steatohepatitis (NASH).

9. The variant human FGF21 protein for use according to any one of the preceding claims, wherein the subject is 18 to 55 years old.

10. The variant human FGF21 protein for use according to any of the preceding claims, wherein the subject has a Body Mass Index (BMI) of 30 to 45kg/m²Is included within the range of (1), wherein for a subject of Asian origin or Asian descent, the ethnic adjustment is > 27.5.

11. The human FGF21 protein variant for use according to any one of the preceding claims, wherein the subject has a triglyceride level in the range of 150-500mg/dL (1.69-5.65mmol/L) when measured before administration of the human FGF21 protein variant.

12. The human FGF21 protein variant for use according to any one of the preceding claims, wherein the human FGF21 variant is a fusion protein comprising a human Fc region fused to a mature human FGF21 protein or a fragment thereof, the mature human FGF21 protein or a fragment thereof comprising one or more mutations selected from the group consisting of: Q55C, R105K, G148C, K150R, P158S, S195A, P199G and G202A numbered according to SEQ ID NO 1.

13. The human FGF21 protein variant for use according to claim 12, wherein the human FGF21 protein variant comprises the amino acid sequence of seq id No. 11.

14. The human FGF21 protein variant for use according to any one of the preceding claims, wherein the human FGF21 protein variant is provided for administration at a dose of at least 100mg, 150mg, 200mg, 250mg, 300mg, 350mg, or 400 mg.

15. The human FGF21 protein variant for use according to any one of the preceding claims, wherein the human FGF21 protein variant is provided for administration at a dose of about 200mg, 210mg, 220mg, 230mg, 240mg, 250mg, 260mg, 270mg, 280mg, 290mg, 300mg, 310mg, 320mg, 330mg, 340mg, or 350mg, optionally wherein the human FGF21 protein variant is provided at a dose of about 250mg or 300 mg.

16. The human FGF21 protein variant for use according to any one of the preceding claims, wherein the human FGF21 protein variant is provided in combination with one or more additional therapeutically active agents.

17. The variant human FGF21 protein for use according to claim 16, wherein the one or more additional therapeutically active agents are selected from the group consisting of: compounds for the treatment of obesity, diuretics, beta-blockers, alpha-blockers, ACE inhibitors, angiotensin II receptor blockers (ARBs), direct renin inhibitors, calcium channel blockers, central agonists, peripheral adrenergic blockers, vasodilators, insulin, alpha-glucosidase inhibitors, biguanides, dopamine agonists, DPP-4 inhibitors, glucagon-like peptides, meglitinides, sodium glucose transporter (SGLT) inhibitors, sulfonylureas, thiazolidinediones, pramlintide, statins, fibrates, aspirin and anticoagulants.

18. The variant human FGF21 protein for use according to claim 17, wherein the sodium glucose transporter (SGLT) inhibitor is selected from dapagliflozin, engagliflozin, canagliflozin, egagliflozin, soagliflozin, togagliflozin, rigagliflozin, luagliflozin, ivagliflozin, begagliflozin, henagliflozin, licogliflozin, and a pharmaceutically acceptable salt of any of these.

19. The human FGF21 protein variant for use according to any one of the preceding claims, wherein the human FGF21 protein variant is provided in a form for subcutaneous administration.

20. The human FGF21 protein variant for use according to any one of the preceding claims, wherein the human FGF21 protein variant is provided for administration monthly or once every 4 weeks, once every 3 weeks, once every 2 weeks, or once a week.